LOCUS       Exported                5935 bp ds-DNA     circular SYN 19-SEP-2018
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5935)
  AUTHORS   Rui Lin
  TITLE     Direct Submission
  JOURNAL   Exported Sep 20, 2018 from SnapGene Viewer 4.2.4
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5935
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     promoter        159..473
                     /label=tight TRE promoter
                     /note="Tet-responsive promoter PTight, consisting of seven 
                     tet operator sequences followed by the minimal CMV 
                     promoter"
     protein_bind    167..185
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    203..221
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    238..256
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    274..292
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    310..328
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    345..363
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    381..399
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    502..535
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GGATACTT). lox2272 sites are compatible with each
                     other, but incompatible with loxP or loxN sites."
     protein_bind    586..619
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     CDS             complement(628..1926)
                     /codon_start=1
                     /product="nuclear-targeted site-specific recombinase"
                     /label=FLPo
                     /note="FLPo is a mouse codon-optimized fusion between the 
                     SV40 nuclear localization signal and a thermostable version
                     of the Saccharomyces cerevisiae site-specific recombinase 
                     FLP.  Recombination occurs at FRT sequences."
                     /translation="MAPKKKRKVMSQFDILCKTPPKVLVRQFVERFERPSGEKIASCAA
                     ELTYLCWMITHNGTAIKRATFMSYNTIISNSLSFDIVNKSLQFKYKTQKATILEASLKK
                     LIPAWEFTIIPYNGQKHQSDITDIVSSLQLQFESSEEADKGNSHSKKMLKALLSEGESI
                     WEITEKILNSFEYTSRFTKTKTLYQFLFLATFINCGRFSDIKNVDPKSFKLVQNKYLGV
                     IIQCLVTETKTSVSRHIYFFSARGRIDPLVYLDEFLRNSEPVLKRVNRTGNSSSNKQEY
                     QLLKDNLVRSYNKALKKNAPYPIFAIKNGPKSHIGRHLMTSFLSMKGLTELTNVVGNWS
                     DKRASAVARTTYTHQITAIPDHYFALVSRYYAYDPISKEMIALKDETNPIEEWQHIEQL
                     KGSAEGSIRYPAWNGIISQEVLDYLSSYINRRI"
     protein_bind    complement(1933..1966)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GGATACTT). lox2272 sites are compatible with each
                     other, but incompatible with loxP or loxN sites."
     protein_bind    complement(2017..2050)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     misc_feature    2065..2673
                     /label=WPRE
     polyA_signal    2682..3158
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     repeat_region   3198..3338
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     rep_origin      3413..3868
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        4150..4254
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4255..5115
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      5286..5874
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca cgcgtctcga gtttactccc tatcagtgat
      181 agagaacgta tgtcgagttt actccctatc agtgatagag aacgatgtcg agtttactcc
      241 ctatcagtga tagagaacgt atgtcgagtt tactccctat cagtgataga gaacgtatgt
      301 cgagtttact ccctatcagt gatagagaac gtatgtcgag tttatcccta tcagtgatag
      361 agaacgtatg tcgagtttac tccctatcag tgatagagaa cgtatgtcga ggtaggcgtg
      421 tacggtggga ggcctatata agcagagctc gtttagtgaa ccgtcagatc gcctggagaa
      481 ttctctagag tcgactccgg aataacttcg tataggatac tttatacgaa gttatgcaga
      541 atggtagctg gattgtagct gctattagca atatgaaacc tcttaataac ttcgtatagc
      601 atacattata cgaagttatg gcgcgcctca gatccgcctg ttgatgtagc tgctcaggta
      661 gtccagcacc tcctggctga tgatgccgtt ccaggcgggg tatctgatgc tgccctcggc
      721 gctgcccttc agctgctcga tgtgctgcca ctcctcgatg gggttggtct cgtccttcag
      781 ggcgatcatc tccttgctga tggggtcgta ggcgtagtac ctggacacca gggcgaagta
      841 gtggtcgggg atggcggtga tctggtgggt gtaggtggtc ctggccacgg cggaggccct
      901 cttgtcgctc cagttgccca ccacgtttgt cagctcggtc aggcccttca tgctcagaaa
      961 gctggtcatc aggtgcctgc cgatgtggct cttagggccg ttcttgatag cgaagatggg
     1021 gtagggggcg ttcttcttca gggccttgtt gtagctgcgc accaggttgt ccttcagcag
     1081 ctggtactcc tgcttgttgc tgctgctgtt gccggtcctg ttcactctct tcagcacggg
     1141 ctcgctgttc ctcaggaact cgtccaggta caccaggggg tcgatcctgc ctctggcgct
     1201 gaaaaagtag atgtgcctgg acacgcttgt cttggtctcg gtcaccaggc actgaatgat
     1261 cacgcccagg tacttgttct gcaccagctt gaagctcttg gggtccacgt tcttgatgtc
     1321 gctgaacctg ccgcagttga tgaatgtggc caggaacagg aactggtaca gggtcttggt
     1381 cttggtgaac ctgctggtgt actcgaagct gttcaggatc ttctcggtga tctcccagat
     1441 gctctcgccc tcggacagca gggccttcag catcttcttg ctgtggctgt tgcccttgtc
     1501 ggcctcctcg ctgctctcga actgcagctg caggctggac acgatgtcgg tgatgtcgct
     1561 ctggtgcttc tggccgttgt aagggatgat ggtgaactcc caggcgggga tcagcttctt
     1621 caggctggcc tccaggatgg tggccttctg ggtcttgtac ttgaactgca ggctcttgtt
     1681 cacgatgtcg aagctcaggc tgttgctgat gatggtgttg tagctcatga aggtggccct
     1741 cttgatggcg gtgccgttgt gggtgatcat ccagcacagg taggtcagct cggcggcaca
     1801 gctggcgatc ttctcgccgc tgggcctctc gaatctctcc acgaactgcc gcaccagcac
     1861 cttggggggg gtcttgcaca ggatgtcgaa ctggctcatc accttcctct tcttcttagg
     1921 agccatgcta gcataacttc gtataaagta tcctatacga agttatttgc cttaacccag
     1981 aaattatcac tgttattctt tagaatggtg caaagaataa cttcgtataa tgtatgctat
     2041 acgaagttat gaattcgata tcaagcttat cgataatcaa cctctggatt acaaaatttg
     2101 tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc
     2161 tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta
     2221 taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt
     2281 ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca
     2341 gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc
     2401 ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt
     2461 gtcggggaaa tcatcgtcct ttccttggct gctcgcctat gttgccacct ggattctgcg
     2521 cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg
     2581 cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat
     2641 ctccctttgg gccgcctccc cgcatcgata ccgagatcta cgggtggcat ccctgtgacc
     2701 cctccccagt gcctctcctg gccctggaag ttgccactcc agtgcccacc agccttgtcc
     2761 taataaaatt aagttgcatc attttgtctg actaggtgtc cttctataat attatggggt
     2821 ggaggggggt ggtatggagc aaggggcaag ttgggaagac aacctgtagg gcctgcgggg
     2881 tctattggga accaagctgg agtgcagtgg cacaatcttg gctcactgca atctccgcct
     2941 cctgggttca agcgattctc ctgcctcagc ctcccgagtt gttgggattc caggcatgca
     3001 tgaccaggct cagctaattt ttgttttttt ggtagagacg gggtttcacc atattggcca
     3061 ggctggtctc caactcctaa tctcaggtga tctacccacc ttggcctccc aaattgctgg
     3121 gattacaggc gtgaaccact gctcccttcc ctgtccttct gattttgtag gtaaccacgt
     3181 gcggaccgag cggccgcagg aacccctagt gatggagttg gccactccct ctctgcgcgc
     3241 tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga cgcccgggct ttgcccgggc
     3301 ggcctcagtg agcgagcgag cgcgcagctg cctgcagggg cgcctgatgc ggtattttct
     3361 ccttacgcat ctgtgcggta tttcacaccg catacgtcaa agcaaccata gtacgcgccc
     3421 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     3481 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     3541 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     3601 cggcacctcg accccaaaaa acttgatttg ggtgatggtt cacgtagtgg gccatcgccc
     3661 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     3721 ttccaaactg gaacaacact caaccctatc tcgggctatt cttttgattt ataagggatt
     3781 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     3841 tttaacaaaa tattaacgtt tacaatttta tggtgcactc tcagtacaat ctgctctgat
     3901 gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct
     3961 tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt
     4021 cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt gatacgccta
     4081 tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg
     4141 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     4201 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     4261 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     4321 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     4381 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     4441 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     4501 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     4561 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     4621 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     4681 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     4741 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     4801 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     4861 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     4921 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     4981 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     5041 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     5101 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     5161 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     5221 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     5281 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     5341 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     5401 ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac
     5461 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     5521 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     5581 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     5641 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     5701 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     5761 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     5821 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     5881 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgt
//
