LOCUS       Exported                3733 bp ds-DNA     circular SYN 17-JUL-2006
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pDONR221-P5P4
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3733)
  AUTHORS   Invitrogen
  TITLE     Direct Submission
  JOURNAL   Exported Jan 17, 2019 from SnapGene Viewer 4.2.11
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..3733
                     /organism="synthetic DNA construct"
                     /note="/dnas_title=KO801_Hygro_P2A"
     misc_feature    complement(268..295)
                     /label=rrnB T2 transcription termination sequence
                     /note="rrnB T2 transcription termination sequence"
     misc_feature    complement(427..470)
                     /label=rrnB T1 transcription termination sequence
                     /note="rrnB T1 transcription termination sequence"
     primer_bind     517..533
                     /label=M13 Forward (-40) primer binding site
                     /note="M13 Forward (-40) primer binding site"
     primer_bind     537..552
                     /label=M13 Forward (-20) primer binding site
                     /note="M13 Forward (-20) primer binding site"
     misc_recomb     570..665
                     /label=attL5
                     /note="attL5 recombination site"
                     /note="recognized as attL5"
     5'UTR           668..733
                     /label=M17374.2
                     /note="Xenopus 5' beta-globin leader (enhances 
                     translation)"
                     /note="M17374.2"
     gene            734..1768
                     /gene="Hygromycin resistance gene"
                     /label=Hygromycin resistance gene
     misc_feature    1772..1780
                     /label=GSG linker
                     /note="GSG linker"
     misc_feature    1781..1837
                     /label=P2A self-cleaving peptide
                     /note="P2A self-cleaving peptide"
     misc_feature    1832..1837
                     /label=G^P cleavage site
                     /note="G^P cleavage site"
     misc_recomb     complement(1845..1943)
                     /label=attL4
                     /note="attL4 recombination site"
                     /note="recognized as attL4"
     misc_feature    join(1855..3733,1..658)
                     /label=nonstandard type: vector
                     /note="/source=pDONR221-P5P4"
                     /note="/type=Donor Vector"
     primer_bind     complement(1998..2014)
                     /label=M13 Reverse primer binding site
                     /note="M13 Reverse primer binding site"
     gene            2127..2936
                     /label=Kanamycin resistance gene
                     /note="Kanamycin resistance gene"
     rep_origin      3057..3730
                     /label=f1 origin
                     /note="f1 origin"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      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 tcgggcccca aataatgatt ttattttgac tgatagtgac
      601 ctgttcgttg caacaaattg atgagcaatg cttttttata atgccaactt tgtatacaaa
      661 agttgtagaa tacaagcttc ttgttctttt tgcagaagct cagaataaac gctcaacttt
      721 ggcagatctg aacatgaaga agcccgaact caccgctacc agcgttgaaa aatttctcat
      781 cgagaagttc gacagtgtga gcgacctgat gcagttgtcg gagggcgaag agagccgagc
      841 cttcagcttc gatgtcggcg gacgcggcta tgtactgcgg gtgaatagct gcgctgatgg
      901 cttctacaaa gaccgctacg tgtaccgcca cttcgccagc gctgcactac ccatccccga
      961 agtgttggac atcggcgagt tcagcgagag cctgacatac tgcatcagta gacgcgccca
     1021 aggcgttact ctccaagacc tccccgaaac agagctgcct gctgtgttac agcctgtcgc
     1081 cgaagctatg gatgctattg ccgccgccga cctcagtcaa accagcggct tcggcccatt
     1141 cgggccccaa ggcatcggcc agtacacaac ctggcgggat ttcatttgcg ccattgctga
     1201 tccccatgtc taccactggc agaccgtgat ggacgacacc gtgtccgcca gcgtagctca
     1261 agccctggac gaactgatgc tgtgggccga agactgtccc gaggtgcgcc acctcgtcca
     1321 tgccgacttc ggcagcaaca acgtcctgac cgacaacggc cgcatcaccg ccgtaatcga
     1381 ctggtccgaa gctatgttcg gggacagtca gtacgaggtg gccaacatct tcttctggcg
     1441 gccctggctg gcttgcatgg agcagcagac tcgctacttc gagcgccggc atcccgagct
     1501 ggccggcagc cctcgtctgc gagcctacat gctgcgcatc ggcctggatc agctctacca
     1561 gagcctcgtg gacggcaact tcgacgatgc tgcctgggct caaggccgct gcgatgccat
     1621 cgtccgcagc ggggccggca ccgtcggtcg cacacaaatc gctcgccgga gcgcagccgt
     1681 atggaccgac ggctgcgtcg aggtgctggc cgacagcggc aaccgccggc ccagtacacg
     1741 accgcgcgct aaggaggtag gtcgagttgc cggatccgga gctactaatt tctctctgtt
     1801 aaagcaggca ggagatgtgg aggagaatcc aggtcctaag gcccacccaa cttttctata
     1861 caaagttggc attataagaa agcattgctt atcaatttgt tgcaacgaac aggtcactat
     1921 cagtcaaaat aaaatcatta tttgccatcc agctgcaggg cggccgcgat atcccctata
     1981 gtgagtcgta ttacatggtc atagctgttt cctggcagct ctggcccgtg tctcaaaatc
     2041 tctgatgtta cattgcacaa gataaaaata tatcatcatg aacaataaaa ctgtctgctt
     2101 acataaacag taatacaagg ggtgttatga gccatattca acgggaaacg tcgaggccgc
     2161 gattaaattc caacatggat gctgatttat atgggtataa atgggctcgc gataatgtcg
     2221 ggcaatcagg tgcgacaatc tatcgcttgt atgggaagcc cgatgcgcca gagttgtttc
     2281 tgaaacatgg caaaggtagc gttgccaatg atgttacaga tgagatggtc agactaaact
     2341 ggctgacgga atttatgcct cttccgacca tcaagcattt tatccgtact cctgatgatg
     2401 catggttact caccactgcg atccccggaa aaacagcatt ccaggtatta gaagaatatc
     2461 ctgattcagg tgaaaatatt gttgatgcgc tggcagtgtt cctgcgccgg ttgcattcga
     2521 ttcctgtttg taattgtcct tttaacagcg atcgcgtatt tcgtctcgct caggcgcaat
     2581 cacgaatgaa taacggtttg gttgatgcga gtgattttga tgacgagcgt aatggctggc
     2641 ctgttgaaca agtctggaaa gaaatgcata aacttttgcc attctcaccg gattcagtcg
     2701 tcactcatgg tgatttctca cttgataacc ttatttttga cgaggggaaa ttaataggtt
     2761 gtattgatgt tggacgagtc ggaatcgcag accgatacca ggatcttgcc atcctatgga
     2821 actgcctcgg tgagttttct ccttcattac agaaacggct ttttcaaaaa tatggtattg
     2881 ataatcctga tatgaataaa ttgcagtttc atttgatgct cgatgagttt ttctaatcag
     2941 aattggttaa ttggttgtaa cactggcaga gcattacgct gacttgacgg gacggcgcaa
     3001 gctcatgacc aaaatccctt aacgtgagtt acgcgtcgtt ccactgagcg tcagaccccg
     3061 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     3121 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     3181 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     3241 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     3301 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     3361 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     3421 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     3481 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     3541 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     3601 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     3661 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     3721 ttgctcacat gtt
//
