LOCUS       Exported                8067 bp ds-DNA     circular SYN 16-DEC-2016
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pSL18-AID-3xHA-DHFR-LoxP-T7
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8067)
  AUTHORS   Trial User
  TITLE     Direct Submission
  JOURNAL   Exported Friday, Dec 16, 2016 from SnapGene 3.2.1
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8067
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     rep_origin      complement(3..458)
                     /direction=LEFT
                     /note="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     600..616
                     /note="M13 fwd"
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             639..668
                     /codon_start=1
                     /note="Linker"
                     /translation="ASKGSGSTQL"
     CDS             669..1352
                     /codon_start=1
                     /gene="Arabidopsis thaliana AXR3"
                     /product="auxin-responsive IAA17 protein"
                     /note="auxin-inducible degron"
                     /note="acts as an auxin-inducible degron (AID) in the 
                     presence of TIR1"
                     /translation="MGSVELNLRETELCLGLPGGDTVAPVTGNKRGFSETVDLKLNLNN
                     EPANKEGSTTHDVVTFDSKEKSACPKDPAKPPAKAQVVGWPPVRSYRKNVMVSCQKSSG
                     GPEAAAFVKVSMDGAPYLRKIDLRMYKSYDELSNALSNMFSSFTMGKHGGEEGMIDFMN
                     ERKLMDLVNSWDYVPSYEDKDGDWMLVGDVPWPMFVDTCKRLRLMKGSDAIGLAPRAME
                     KCKSRA"
     CDS             1389..1415
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /note="HA"
                     /translation="YPYDVPDYA"
     CDS             1419..1445
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /note="HA"
                     /translation="YPYDVPDYA"
     CDS             1446..1472
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /note="HA"
                     /translation="YPYDVPDYA"
     misc_feature    1482..1901
                     /note="HX 3'UTR"
     protein_bind    1951..1984
                     /bound_moiety="Cre recombinase"
                     /note="loxP"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     misc_feature    1985..2977
                     /note="DHFR 5'UTR"
     misc_feature    2978..4807
                     /note="DHFR"
     protein_bind    4827..4860
                     /bound_moiety="Cre recombinase"
                     /note="loxP"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     misc_feature    4861..5823
                     /note="DHFR 3'UTR"
     promoter        complement(5860..5878)
                     /note="T3 promoter"
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5899..5915)
                     /note="M13 rev"
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(5916..5934)
                     /note="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5942..5958
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="lac operator"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(5966..5996)
                     /note="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    6011..6032
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding site"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(6320..6908)
                     /direction=LEFT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7079..7939)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /note="AmpR"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(7940..8044)
                     /gene="bla"
                     /note="AmpR promoter"
ORIGIN
        1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
       61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
      121 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
      181 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
      241 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
      301 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
      361 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
      421 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg
      481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
      541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
      601 taaaacgacg gccagtgagc gaattgggta ccgggcccgc tagcaagggc tcgggctcga
      661 cccagctgat gggcagtgtc gagctgaatc tgagggagac tgagctgtgt cttggtcttc
      721 ccggtggaga tacagtggct ccggtaaccg gaaacaagag agggttctca gagacggttg
      781 atctgaagct aaatctgaat aatgagcctg caaacaagga aggatctacg actcatgacg
      841 tcgtgacttt tgattccaag gagaagagtg cttgtcctaa agatccagcc aaacctccgg
      901 ccaaggcaca agttgtggga tggccaccgg tgagatcata ccggaagaac gtgatggttt
      961 cctgccaaaa atcaagcggt ggcccggagg cggcggcgtt cgtgaaggta tcaatggacg
     1021 gagcaccgta cttgaggaaa atcgatttga ggatgtataa aagctacgat gagctttcta
     1081 atgctttgtc caacatgttc agctctttta ccatgggcaa acatggagga gaagaaggaa
     1141 tgatagactt catgaatgag aggaaattga tggatttggt gaatagctgg gactatgttc
     1201 cctcttatga agacaaagac ggtgattgga tgctcgtcgg cgacgttcct tggccaatgt
     1261 tcgtcgatac atgcaagcgt ttacgtctca tgaaaggatc ggatgccatt ggtctcgctc
     1321 cgagggcgat ggagaagtgc aagagcagag ctgggcgcgc cggctctggc ggcggcggcg
     1381 gcggctctta cccgtacgac gtcccggact acgctggcta tccctatgat gtgcccgatt
     1441 atgcgtatcc ttacgatgtt ccagattatg cctaacccgg gcatatgtag aaaagttgta
     1501 acgttagtaa acgtaacaca cgggtataat gccatttaat ggcgccgtat atttttcact
     1561 ccccctctcc gataggagga aaaagacgat ccgtctgcga atgtatatta catcgaccgg
     1621 ccatagtacg agaagtggtg ccggtattgt tctgtccggg ggatgtacca tcctattttt
     1681 tgtatatgtc gttcttgtcg gggagcaaca gcccatcttc cggctattgt ctcgtatcag
     1741 ttgtgaaaca gactgcgttt tgccgaaaat gagtcctatg tccgatggct atttgtaccg
     1801 cttcgtacca tttcgcttgt cagctgtaat caaccgaatt catttggcga gacatgattc
     1861 cgattaaatt ctttactgcg cgtgctcgga tccactagtt ctactcgagg gtcgacggta
     1921 tcgattctag agaattcgag ctcggtaccc ataacttcgt atagcataca ttatacgaag
     1981 ttataagctt cgccaggctg taaatcccgt gagtcgtcct cacaaatcat caagcaggtg
     2041 tcctcaggga gactgcctga ctgagttatg ctaattcctt tctactttgg cgtggtcacg
     2101 ggggcgcgcc ggatccttaa ttaagtctag catgtcattc gattttcacc ccccgcgtag
     2161 ttcctgtgtg tcattcgttg tcgagacaac tctgtcccgc cccggtgctg ttccatatgc
     2221 gtgactttcc cgcaattttt tcagactttc aggaaagaca ggctccggaa cgatctcgtc
     2281 catgactggt aaatccacga caccgcaatg gcccccagca cctctatctc tcgtgccagg
     2341 ggactaacgt tgtatgcgtc tgcgtcttgt ctttttgcat tcgctttcca aaaaagagag
     2401 ccatccgttc ccccgcacat tcaacgccgc gagtgcggtt tttgtctttt ttgagtggta
     2461 ggacgctttt catgcgcgaa ctacgtggac attaagttcc attctctttt tcgacagcac
     2521 gaaaccttgc attcaaaccc gcccgcggaa gatccgatct tgctgctgtt cgcagtccca
     2581 gtagcgtcct gtcggccgcg ccgtctctgt tggtgggcag ccgctacacc tgttatctga
     2641 ctgccgtgcg cgaaaatgac gccatttttg ggaaaatcgg ggaacttcat tctttaaaag
     2701 tatgcggagg tttccttttt cttctgttcg tttctttttc tcgggtttga taaccgtgtt
     2761 cgatgtaagc actttccgtc tctcctccgt gctttgttcg acatcgagac caggtgtgca
     2821 gatccttcgc ttgtcgatcc ggagacgcgt gtctcgtaga accttttcat tttaccacac
     2881 ggcagtgcgg agcactgctc tgagtgcagc agggacgggt gaagtttcgc tttagtagtg
     2941 cgtttctgct ctacggggcg ttgtcgtgtc tgggaagatg cagaaaccgg tgtgtctggt
     3001 cgtcgcgatg acccccaaga ggggcatcgg catcaacaac ggcctcccgt ggccccactt
     3061 gaccacagat ttcaaacact ttcgtcgtgt gacaaaaacg acgcccgaag aagccagtcg
     3121 cctgaacggg tggcttccca ggaaatttgc aaagacgggc gactctggac ttccctctcc
     3181 atcagtcggc aagagattca acgccgttgt catgggacgg aaaaactggg aaagcatgcc
     3241 tcgaaagttt agacccctcg tggacagatt gaacatcgtc gtttcctctt ccctcaaaga
     3301 agaagacatt gcggcggaga agcctcaagc tgaaggccag cagcgcgtcc gagtctgtgc
     3361 ttcactccca gcagctctca gccttctgga ggaagagtac aaggattctg tcgaccagat
     3421 ttttgtcgtg ggaggagcgg gactgtacga ggcagcgctg tctctgggcg ttgcctctca
     3481 cctgtacatc acgcgtgtag cccgcgagtt tccgtgcgac gttttcttcc ctgcgttccc
     3541 cggagatgac attctttcaa acaaatcaac tgctgcgcag gctgcagctc ctgccgagtc
     3601 tgtgttcgtt cccttttgtc cggagctcgg aagagagaag gacaatgaag cgacgtatcg
     3661 acccatcttc atttccaaga ccttctcaga caacggggta ccctacgact ccgtggttct
     3721 cgagaagaga aggaagactg acgacgcagc cactgcggaa ccgagcaacg caatgagctc
     3781 cttgacgtcc acgagggaga caactcccgt gcacgggttg caggctcctt cttcggccgc
     3841 agccattgcc ccggtgttgg cgtggatgga cgaagaagac cggaaaaaac gcgagcaaaa
     3901 ggaactgatt cgggccgttc cgcatgttca ctttagaggc catgaagaat tccagtacct
     3961 tgatctcatt gccgacatta ttaacaatgg aaggacaatg gatgaccgaa cgggcgttgg
     4021 tgtcatctcc aaattcggct gcactatgcg ctactcgctg gatcaggcct ttccacttct
     4081 caccacaaag cgtgtgttct ggaaaggggt cctcgaagag ttgctgtggt tcattcgcgg
     4141 cgacacgaac gcaaaccatc tttctgagaa gggcgtgaag atctgggaca agaatgtgac
     4201 acgcgagttc ctcgattcgc gcaatctccc ccaccgagag gtcggagaca tcggcccggg
     4261 ctacggcttc cagtggagac acttcggcgc ggcatacaaa gacatgcaca cagactacac
     4321 agggcagggc gtcgaccagc tgaagaatgt gatccagatg ctgagaacga atccaacaga
     4381 tcgtcgcatg ctcatgactg cctggaatcc tgcagcgctg gacgaaatgg cgctgccgcc
     4441 ttgtcacttg ttgtgccagt tctacgtgaa cgaccagaag gagctgtcgt gcatcatgta
     4501 tcagcggtcg tgcgatgtcg gcctcggcgt ccccttcaac atcgcttcct attcgctttt
     4561 gacgctcatg gttgcacacg tctgcaacct aaaacctaag gagttcattc acttcatggg
     4621 gaacacgcat gtctacacga accatgtcga ggctttaaaa gagcagctgc ggagagaacc
     4681 gagaccgttc cccattgtga acatcctcaa caaggaacgc atcaaggaaa tcgacgattt
     4741 caccgccgag gattttgagg tcgtgggcta cgtcccgcac ggacgaatcc agatggagat
     4801 ggctgtcggt tacaagtaag ctgagcataa cttcgtatag catacattat acgaagttat
     4861 cggaaataca gaagctgccc gtctctcgtt ttcctctctt ttcggaggga tcagggagag
     4921 tgcctcgggt cggagagagc tgacgagggg gtgccagaga cccctgtgtc ctttatcgaa
     4981 gaaaagggat gactcttcat gtggcatttc acacagtctc acctcgcctt gttttctttt
     5041 tgtcaatcag aacgaaagcg agttgcgggt gacgcagatg tgcgtgtatc cactcgtgaa
     5101 tgcgttatcg ttctgtatgc cgctagagtg ctggactgtt gctgtctgcc cacgacagca
     5161 gacaactttc cttctatgca cttgcaggat ggtgcagcgc aaacgacgga gagaaaggag
     5221 caccctctca gtttccctac gatgtgctgt cagtttcgac tcttcaccgc gaacgattgg
     5281 cgatacgtct ctgttgactt gttaggctcc gaccacgaag ctcccttaac taaataagcc
     5341 gcgacaccta agtgtacacc atttgcagat cgataatctg cgaccgctga atccgtccag
     5401 atcagtaaaa ccgcaccacc taagtgtaaa ccttgtttag gtcgataaaa tgctaccaac
     5461 ccccacccac aatcgagcct tgagcgtttc tgcgcacgcg ttggcctacg tgacttgctg
     5521 atgcctgcct ctggccattc atgccagtca gtgcgcataa aaatgtggac acagtcggtt
     5581 gacaagtgtt ctggcaggct acagtgacac ggggaaatcg tttctgagag agcaccaaaa
     5641 ccttttagct gtcaccatct tcttcgcttg ctacgttgtc tgatttgtcg tttttcctct
     5701 cggtgtgaag cgtttttcgt aacgtcaatc gacccaggac actgcttcca gctgtctgtg
     5761 ccgtcctgtc gtgaccagtt tgtcgccgcg actttatgag ctttttcctg ttgagaaagc
     5821 ggtgcggccg ccaccgcggt ggagctccag cttttgttcc ctttagtgag ggttaattgc
     5881 gcgcttggcg taatcatggt catagctgtt tcctgcctat agtgagtcgt attatgtgaa
     5941 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     6001 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     6061 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     6121 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     6181 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     6241 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     6301 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     6361 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     6421 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     6481 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     6541 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     6601 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     6661 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     6721 agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg
     6781 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     6841 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     6901 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     6961 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     7021 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     7081 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     7141 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     7201 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     7261 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     7321 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     7381 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     7441 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     7501 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     7561 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     7621 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     7681 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     7741 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     7801 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     7861 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     7921 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     7981 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     8041 cgcgcacatt tccccgaaaa gtgccac
//
