LOCUS       Exported                5329 bp ds-DNA     circular SYN 17-JUL-2006
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pDONR221-P3P2
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5329)
  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..5329
                     /organism="synthetic DNA construct"
                     /note="/dnas_title=KO701_HA_PolyA_EF1a_TETa_PolyA"
     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=attL3
                     /note="attL3 recombination site"
                     /note="recognized as attL3"
     gene            686..715
                     /label=HA epitope tag
                     /note="HA epitope tag"
     polyA_signal    733..932
                     /label=SV40 polyA
                     /note="SV40 polyA"
     promoter        939..2122
                     /note="EF1a promoter"
     promoter        2139..2158
                     /note="T7 promoter"
     gene            2199..2945
                     /label=Tet-On Advanced
                     /note="Tet-On Advanced"
     polyA_signal    2968..3410
                     /label=SV40 polyA
                     /note="SV40 polyA"
     misc_recomb     complement(3441..3539)
                     /label=attL2
                     /note="attL2 recombination site"
                     /note="recognized as attL2"
     misc_feature    join(3451..5329,1..658)
                     /label=nonstandard type: vector
                     /note="/source=pDONR221-P3P2"
                     /note="/type=Donor Vector"
     primer_bind     complement(3594..3610)
                     /label=M13 Reverse primer binding site
                     /note="M13 Reverse primer binding site"
     gene            3723..4532
                     /label=Kanamycin resistance gene
                     /note="Kanamycin resistance gene"
     rep_origin      4653..5326
                     /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 tgtataataa
      661 agttgagatc aacaaagctg gatcctaccc atacgatgtg ccagattacg cttgatttaa
      721 atagataact gactagagga tcataatcag ccataccaca tttgtagagg ttttacttgc
      781 tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt
      841 tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
      901 cacaaataaa gcattttttt cactgcctcg agtctagacg tgaggctccg gtgcccgtca
      961 gtgggcagag cgcacatcgc ccacagtccc cgagaagttg gggggagggg tcggcaattg
     1021 aaccggtgcc tagagaaggt ggcgcggggt aaactgggaa agtgatgtcg tgtactggct
     1081 ccgccttttt cccgagggtg ggggagaacc gtatataagt gcagtagtcg ccgtgaacgt
     1141 tctttttcgc aacgggtttg ccgccagaac acaggtaagt gccgtgtgtg gttcccgcgg
     1201 gcctggcctc tttacgggtt atggcccttg cgtgccttga attacttcca cctggctgca
     1261 gtacgtgatt cttgatcccg agcttcgggt tggaagtggg tgggagagtt cgaggccttg
     1321 cgcttaagga gccccttcgc ctcgtgcttg agttgaggcc tggcctgggc gctggggccg
     1381 ccgcgtgcga atctggtggc accttcgcgc ctgtctcgct gctttcgata agtctctagc
     1441 catttaaaat ttttgatgac ctgctgcgac gctttttttc tggcaagata gtcttgtaaa
     1501 tgcgggccaa gatctgcaca ctggtatttc ggtttttggg gccgcgggcg gcgacggggc
     1561 ccgtgcgtcc cagcgcacat gttcggcgag gcggggcctg cgagcgcggc caccgagaat
     1621 cggacggggg tagtctcaag ctggccggcc tgctctggtg cctggcctcg cgccgccgtg
     1681 tatcgccccg ccctgggcgg caaggctggc ccggtcggca ccagttgcgt gagcggaaag
     1741 atggccgctt cccggccctg ctgcagggag ctcaaaatgg aggacgcggc gctcgggaga
     1801 gcgggcgggt gagtcaccca cacaaaggaa aagggccttt ccgtcctcag ccgtcgcttc
     1861 atgtgactcc acggagtacc gggcgccgtc caggcacctc gattagttct cgagcttttg
     1921 gagtacgtcg tctttaggtt ggggggaggg gttttatgcg atggagtttc cccacactga
     1981 gtgggtggag actgaagtta ggccagcttg gcacttgatg taattctcct tggaatttgc
     2041 cctttttgag tttggatctt ggttcattct caagcctcag acagtggttc aaagtttttt
     2101 tcttccattt caggtgtcgt gaggaattag cttggtacta atacgactca ctatagggag
     2161 acccaagctg gctaggtgaa ttcgactggc gcgccaccat gtccagactg gacaagagca
     2221 aagtcataaa cggcgctctg gaattactca atggagtcgg tatcgaaggc ctgacgacaa
     2281 ggaaactcgc tcaaaagctg ggagttgagc agcctaccct gtactggcac gtgaagaaca
     2341 agcgggccct gctcgatgcc ctgccaatcg agatgctgga caggcatcat acccacttct
     2401 gccccctgga aggcgagtca tggcaagact ttctgcggaa caacgccaag tcattccgct
     2461 gtgctctcct ctcacatcgc gacggggcta aagtgcatct cggcacccgc ccaacagaga
     2521 aacagtacga aaccctggaa aatcagctcg cgttcctgtg tcagcaaggc ttctccctgg
     2581 agaacgcact gtacgctctg tccgccgtgg gccactttac actgggctgc gtattggagg
     2641 aacaggagca tcaagtagca aaagaggaaa gagagacacc taccaccgat tctatgcccc
     2701 cacttctgag acaagcaatt gagctgttcg accggcaggg agccgaacct gccttccttt
     2761 tcggcctgga actaatcata tgtggcctgg agaaacagct aaagtgcgaa agcggcgggc
     2821 cggccgacgc ccttgacgat tttgacttag acatgctccc agccgatgcc cttgacgact
     2881 ttgaccttga tatgctgcct gctgacgctc ttgacgattt tgaccttgac atgctccccg
     2941 ggtaactaag taagaatcca gacatgataa gatacattga tgagtttgga caaaccacaa
     3001 ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg
     3061 taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc
     3121 aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta aaacctctac aaatgtggta
     3181 tggctgatta tgatcctgca agcctcgtcg tcctggccgg accacgctat ctgtgcaagg
     3241 tccccggccc cggacgcgcg ctccatgagc agagcgcccg ccgccgaggc gaagactcgg
     3301 gcggcgccct gcccgtccca ccaggtcaac aggcggtaac cggcctcttc atcgggaatg
     3361 cgcgcgacct tcagcatcgc cggcatgtcc ccctggcgga cgggaagtat ccagctcgac
     3421 caggcttgtc gactcgaaga acccagcttt cttgtacaaa gttggcatta taagaaagca
     3481 ttgcttatca atttgttgca acgaacaggt cactatcagt caaaataaaa tcattatttg
     3541 ccatccagct gcagggcggc cgcgatatcc cctatagtga gtcgtattac atggtcatag
     3601 ctgtttcctg gcagctctgg cccgtgtctc aaaatctctg atgttacatt gcacaagata
     3661 aaaatatatc atcatgaaca ataaaactgt ctgcttacat aaacagtaat acaaggggtg
     3721 ttatgagcca tattcaacgg gaaacgtcga ggccgcgatt aaattccaac atggatgctg
     3781 atttatatgg gtataaatgg gctcgcgata atgtcgggca atcaggtgcg acaatctatc
     3841 gcttgtatgg gaagcccgat gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg
     3901 ccaatgatgt tacagatgag atggtcagac taaactggct gacggaattt atgcctcttc
     3961 cgaccatcaa gcattttatc cgtactcctg atgatgcatg gttactcacc actgcgatcc
     4021 ccggaaaaac agcattccag gtattagaag aatatcctga ttcaggtgaa aatattgttg
     4081 atgcgctggc agtgttcctg cgccggttgc attcgattcc tgtttgtaat tgtcctttta
     4141 acagcgatcg cgtatttcgt ctcgctcagg cgcaatcacg aatgaataac ggtttggttg
     4201 atgcgagtga ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa
     4261 tgcataaact tttgccattc tcaccggatt cagtcgtcac tcatggtgat ttctcacttg
     4321 ataaccttat ttttgacgag gggaaattaa taggttgtat tgatgttgga cgagtcggaa
     4381 tcgcagaccg ataccaggat cttgccatcc tatggaactg cctcggtgag ttttctcctt
     4441 cattacagaa acggcttttt caaaaatatg gtattgataa tcctgatatg aataaattgc
     4501 agtttcattt gatgctcgat gagtttttct aatcagaatt ggttaattgg ttgtaacact
     4561 ggcagagcat tacgctgact tgacgggacg gcgcaagctc atgaccaaaa tcccttaacg
     4621 tgagttacgc gtcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     4681 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     4741 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     4801 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     4861 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     4921 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     4981 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     5041 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     5101 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     5161 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     5221 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     5281 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgtt
//
