LOCUS       Exported                7350 bp ds-DNA     circular SYN 01-MAY-2019
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    pCR8_GW_TOPO_Cdk12_isoform1NFH
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7350)
  AUTHORS   Sara Dubbury
  TITLE     Direct Submission
  JOURNAL   Exported Wednesday, May 1, 2019 from SnapGene 3.3.4
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7350
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     terminator      268..295
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
                     /note="color: #ffffff"
     terminator      387..473
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
                     /note="color: #ffffff"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /note="color: #a020f0; direction: RIGHT"
     protein_bind    569..668
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
                     /note="color: #31849b; direction: RIGHT"
     misc_feature    681..5215
                     /label=TOPO_Insertion_Site
                     /note="color: #800000"
     misc_feature    683..777
                     /note="SJD102"
                     /note="color: #50ff50"
     misc_feature    704..757
                     /note="FLAG- HA-"
                     /note="color: #0080ff"
     CDS             707..730
                     /codon_start=1
                     /label=FLAG Epitope
                     /note="color: #993366"
                     /translation="DYKDDDDK"
     CDS             731..757
                     /codon_start=1
                     /label=HA epitope
                     /note="color: #993366"
                     /translation="YPYDVPDYA"
     primer_bind     734..753
                     /label=Cdk12_FlagHA_F1
                     /note="color: #a020f0; direction: RIGHT"
     misc_feature    737..755
                     /note="SJD324"
                     /note="color: #50ff50"
     misc_feature    758..1797
                     /note="exon1"
                     /note="color: #0080ff"
     misc_feature    complement(874..893)
                     /note="SJD325"
                     /note="color: #50ff50; direction: LEFT"
     primer_bind     complement(879..898)
                     /label=Cdk12_ex1_R1
                     /note="color: #a020f0; direction: LEFT"
     primer_bind     complement(893..912)
                     /label=Cdk12_ex1_R2
                     /note="color: #a020f0; direction: LEFT"
     misc_feature    1798..2673
                     /note="exon2"
                     /note="color: #0080ff"
     misc_feature    2674..2850
                     /note="exon3"
                     /note="color: #0080ff"
     misc_feature    2801..2820
                     /note="SJD326"
                     /note="color: #50ff50"
     misc_feature    2851..2990
                     /note="exon4"
                     /note="color: #0080ff"
     misc_feature    complement(2934..2953)
                     /note="SJD327"
                     /note="color: #50ff50; direction: LEFT"
     misc_feature    2939..2958
                     /note="SJD328"
                     /note="color: #50ff50"
     misc_feature    2991..3161
                     /note="exon5"
                     /note="color: #0080ff"
     misc_feature    complement(3029..3048)
                     /note="SJD329"
                     /note="color: #50ff50; direction: LEFT"
     misc_feature    3162..3351
                     /note="exon6"
                     /note="color: #0080ff"
     misc_feature    3352..3408
                     /note="exon7"
                     /note="color: #0080ff"
     misc_feature    3409..3510
                     /note="exon8"
                     /note="color: #0080ff"
     misc_feature    3511..3588
                     /note="exon9"
                     /note="color: #0080ff"
     misc_feature    3589..3705
                     /note="exon10"
                     /note="color: #0080ff"
     misc_feature    3706..3837
                     /note="exon11"
                     /note="color: #0080ff"
     misc_feature    3838..4049
                     /note="exon12"
                     /note="color: #0080ff"
     misc_feature    4050..4502
                     /note="exon13"
                     /note="color: #0080ff"
     misc_feature    4503..5209
                     /note="exon14"
                     /note="color: #0080ff"
     misc_feature    complement(5189..5215)
                     /note="SJD35"
                     /note="color: #50ff50; direction: LEFT"
     protein_bind    complement(5229..5328)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
                     /note="color: #31849b; direction: LEFT"
     promoter        complement(5346..5364)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /note="color: #ffffff; direction: LEFT"
     primer_bind     complement(5369..5385)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /note="color: #a020f0; direction: LEFT"
     CDS             5816..6607
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /note="color: #ccffcc"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     rep_origin      6700..7288
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
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 tgtacaaaaa
      661 agcaggctcc gaattcgccc ttactactga gctcgccgcc accatggact acaaggacga
      721 cgatgacaag tacccttatg acgtgcccga ttacgctccc aattccgaga gacatggggg
      781 caagaaggac gggagcggag gagcttctgg aacttcgcag ccgtcatcgg gaggtggcag
      841 ctccaacagc agggagcgtc accgcttggt gtcgaagcac aagcggcata agtccaagca
      901 ctccaaagac gtggggctgg tgacccccga agcggcatct ttgggtacca taatcaaacc
      961 actggtggag tacgatgaca tcagctctga ttcagacacc ttctccgatg acacggcctt
     1021 caaatcagac cggagggaga acgaggaacg tcggggaacg gatcggagcg atcgcctgca
     1081 ccgacatcgt caccaccagc accggcggtc ccgagacttg ctaaaaacta aacagacgga
     1141 aaaggaaaaa aatcaggaag tctccaaatc tggatctatg aaggaccggg tatcgggcag
     1201 ttccaaacgg tccgtggagg ggagtgatga ttatgggaag gcccagctat ccaaaagcgg
     1261 cagcaaggaa tccaggtcgt ccaaaatgca caaggagaag acccgcaaag agcgagagtt
     1321 aaagtctgga tacaaggacc ggagtaaaag tcatcggaaa agggaaacac ccaaaagtta
     1381 caaaaccgtg gctagcccta aacggagatc caggagtccc cataggaaat ggtctgacag
     1441 ttccaagcaa gatgacagcc cttccggagc ttcttatggc caagactacg atcttagccc
     1501 cccaaggtct cacacttcta gcaactatga ctcctacaag aagagtcctg gaagtacctc
     1561 aagaaggcag tcaatcagcc caccttacaa agagccttct gcttaccagt ccagcactcg
     1621 gtcacccagt ccttacagcc gacgacagag gtctgtgagt ccctatagcc ggagacggtc
     1681 ctccagctat gaaaggagcg gctcttacag cgggagatca cccagcccct atggccgaag
     1741 gcgatcaagc agccctttcc tgagcaagag gtctctgagt cggagtccac tccccagtag
     1801 gaaatccatg aagtccagaa gtagaagtcc tgcatattca agacactcat cttctcatag
     1861 taaaaagaag cgatccgggt cacgcagtcg tcattccagt atctcacctg tcaggcttcc
     1921 attgaattcc agcctgggag ctgaactcag tagaaaaaag aaggaaagag cagcagctgc
     1981 tgcagcagca aaaatggatg gaaaagagtc caagagttca cctataattt tgcctaaaaa
     2041 agagaaactt gaggtgaagg agtcagggtt agagtctaaa aagttaccca gaagtataaa
     2101 atcagaaaaa tcgaccccag atactgaact ggttactgta gcacattcaa acccagaggt
     2161 aaaacattgt ttagacacag ggaaggttag gttggatgag aacttgcaga agcatcctgc
     2221 taaggatttg aaagcacagg gaacaaagga cgttaaacct gtagcaccga aagaggtgat
     2281 tgttacttca aaggagacag agacatcaga aaaggagacc cttccacctc ttcccacaat
     2341 tacttctcca ccccctttac cagctactac ccctccacct cagacacccc ctttgccacc
     2401 tttgcctcca ctaccagcta ttccgctgca gccacctctg cctcctcccc aaccaccatt
     2461 tagtcaagtt cctgtttcaa gtacttcaat tttaccctct tctcctcacc caaggacatc
     2521 tactctatcc tctcagacaa attctcagcc ccctgtacag gtttctatga agactcaagt
     2581 atctataaca gctgctattc cacatctgaa gacttcaaca ttgcctcctc tgcccctccc
     2641 tcccctatta cctggagatg atgacatgga tagtccaaaa gaaacacttc cttcaaagcc
     2701 tgcaaagaaa gagaaggaac agaggactcg ccacttgctt acagacttgc ctcttcctcc
     2761 tgagctacca ggaggagatc catcgcctcc agattctcca gagccaaagg caattacacc
     2821 acctcaacaa ccatataaaa agagaccaaa aatttgttgt ccacgttatg gagaaagaag
     2881 acaaacagaa agtgattggg ggaagcgctg tgtggacaag tttgacatta ttgggattat
     2941 tggagagggg acctatggcc aagtatataa agccaaggac aaagacacag gagaactagt
     3001 agctctgaag aaggttcggc tggacaatga gaaagagggc ttcccaatca cagccatccg
     3061 ggagatcaag attcttcgtc aattagttca ccagagtgtt gtaaacatga aggaaattgt
     3121 cacagacaaa caagatgcac tggatttcaa gaaggacaaa ggtgcctttt accttgtatt
     3181 tgaatatatg gaccatgact taatgggact gcttgaatca ggtttggtgc acttttctga
     3241 ggaccatatc aagtcattta tgaaacagct aatggaagga ctggattact gtcacaaaaa
     3301 gaatttcctc catcgggata ttaaatgttc taacattttg ctgaataaca gcgggcaaat
     3361 caaactggca gattttggac ttgctcggct ctataactct gaagagagtc gcccttacac
     3421 aaacaaagtc atcactcttt ggtatcgacc tccagagctt cttcttggag aggaaagata
     3481 cacaccagcc attgatgttt ggagctgtgg gtgcatcctt ggagaactgt tcacaaagaa
     3541 acctattttt caagccaatt tagaactggc tcagctagaa cttatcagtc gtctctgtgg
     3601 tagtccttgt ccagcagtgt ggcctgatgt tatcaagctg ccctacttca acaccatgaa
     3661 accgaagaag caatacagga gacgcctaag agaagaattc tctttcattc cttcagcggc
     3721 acttgatcta ttggaccaca tgctgacact ggatcctagc aagaggtgca cagctgaaca
     3781 gaccctacag agtgactttc ttaaagatgt ggaactcagc aaaatggcac ctccagacct
     3841 acctcactgg caggattgcc atgaattgtg gagtaagaaa cgtcgacggc agcgacagag
     3901 tggtattgtg atagaagatc cacctccgtc caaagcttct agaaaagaaa ctacctcagg
     3961 gacaacagct gagcctgtga aaaacaatag cccagcacca cctcagcctg ctcctgtcaa
     4021 ggcagagcct ggtccagggg atgcagtagg ccttggtgac atcacacagc agttgaatca
     4081 aagtgaattg gcagtgttat taaacctgct tcagagccaa actgacctga gcatccctca
     4141 gatggcacag ctgcttaata tccactccaa tccagagatg caacagcagc ttgaagcctt
     4201 gaatcagtct attagtgcac tgactgaagc cagttcccag cagcaggact cagaatccat
     4261 agcccctgaa gaatcattga aggaggtacc ttctgtacct gtggtcctgc ctcctgctga
     4321 acagacaact cctgaagctt caaacacacc agctgacatg cagaatgtgt tggcagttct
     4381 cttgagtcag ctgatgaaaa cccaagagcc agcaggtaac ctggaggaaa acaccaatga
     4441 caagaatagt gggccacagg ggccccgaag aactcctaca atgccacagg aggaggcagc
     4501 agcatgtcct cctcacattc ttccaccaga gaagaggccc cctgagcccc ctggacctcc
     4561 accgccgcca cctccacccc ctctggttga aggcgatctt tccagcgccc cccaggagtt
     4621 gaatcccgcc gtgacagccg ccttgctgca acttttatcc cagcctgaag cagagcctcc
     4681 tggccacctg ccacatgagc accaggcctt gagaccaatg gaatactcca cccgatccca
     4741 tccaaacagg acttacggaa atactgatgg gcctgagaca gggttcagtt ccgctgacac
     4801 tgatgaacgc agttctggtc cagccttgac agaatctttg gttcagaccc cggtgaagaa
     4861 caggaccttc tcaggctctg tgagccacct tggggagtcc aacagctacc agggcacagg
     4921 gtcagtgcag ttcccagggg accaggacct ccgttttacc agggttccct tagcattaca
     4981 ctcagtggtt gggcaaccat tcctcaagtc tgagggaaat agcaactctg tggtacatgc
     5041 agagaccaaa ttgcaaaact atggggagct gggaccggga accactgggg ccaacagctc
     5101 aggaacaacg cttcagtggg ggggcccagc tcagtcttat ggaaaaccct acaggggggc
     5161 tgcaagagtc ctaccacgag ggggaagagg gagaggagtt ccttattaac tcgagaaggg
     5221 cgaattcgac ccagctttct tgtacaaagt tggcattata aaaaataatt gctcatcaat
     5281 ttgttgcaac gaacaggtca ctatcagtca aaataaaatc attatttgcc atccagctga
     5341 tatcccctat agtgagtcgt attacatggt catagctgtt tcctggcagc tctggcccgt
     5401 gtctcaaaat ctctgatgtt acattgcaca agataaaaat atatcatcat gcctcctcta
     5461 gaccagccag gacagaaatg cctcgacttc gctgctgccc aaggttgccg ggtgacgcac
     5521 accgtggaaa cggatgaagg cacgaaccca gtggacataa gcctgttcgg ttcgtaagct
     5581 gtaatgcaag tagcgtatgc gctcacgcaa ctggtccaga accttgaccg aacgcagcgg
     5641 tggtaacggc gcagtggcgg ttttcatggc ttgttatgac tgtttttttg gggtacagtc
     5701 tatgcctcgg gcatccaagc agcaagcgcg ttacgccgtg ggtcgatgtt tgatgttatg
     5761 gagcagcaac gatgttacgc agcagggcag tcgccctaaa acaaagttaa acatcatgag
     5821 ggaagcggtg atcgccgaag tatcgactca actatcagag gtagttggcg tcatcgagcg
     5881 ccatctcgaa ccgacgttgc tggccgtaca tttgtacggc tccgcagtgg atggcggcct
     5941 gaagccacac agtgatattg atttgctggt tacggtgacc gtaaggcttg atgaaacaac
     6001 gcggcgagct ttgatcaacg accttttgga aacttcggct tcccctggag agagcgagat
     6061 tctccgcgct gtagaagtca ccattgttgt gcacgacgac atcattccgt ggcgttatcc
     6121 agctaagcgc gaactgcaat ttggagaatg gcagcgcaat gacattcttg caggtatctt
     6181 cgagccagcc acgatcgaca ttgatctggc tatcttgctg acaaaagcaa gagaacatag
     6241 cgttgccttg gtaggtccag cggcggagga actctttgat ccggttcctg aacaggatct
     6301 atttgaggcg ctaaatgaaa ccttaacgct atggaactcg ccgcccgact gggctggcga
     6361 tgagcgaaat gtagtgctta cgttgtcccg catttggtac agcgcagtaa ccggcaaaat
     6421 cgcgccgaag gatgtcgctg ccgactgggc aatggagcgc ctgccggccc agtatcagcc
     6481 cgtcatactt gaagctagac aggcttatct tggacaagaa gaagatcgct tggcctcgcg
     6541 cgcagatcag ttggaagaat ttgtccacta cgtgaaaggc gagatcacca aggtagtcgg
     6601 caaataaccc tcgagccacc catgaccaaa atcccttaac gtgagttacg cgtcgttcca
     6661 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     6721 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     6781 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     6841 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     6901 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     6961 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     7021 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     7081 acagcgtgag cattgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     7141 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     7201 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     7261 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     7321 ggccttttgc tggccttttg ctcacatgtt
//
