LOCUS       Exported                7928 bp ds-DNA     circular SYN 19-MAR-2018
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7928)
  AUTHORS   mei
  TITLE     Direct Submission
  JOURNAL   Exported Mar 27, 2018 from SnapGene 4.1.7
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7928
                     /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"
     misc_feature    156..681
                     /label=per2 promoter
     protein_bind    700..733
                     /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    784..817
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     CDS             complement(823..942)
                     /codon_start=1
                     /product="PEST degradation sequence from mouse ornithine 
                     decarboxylase"
                     /label=hPEST
                     /note="human codon-optimized"
                     /translation="SHGFPPEVEEQAAGTLPMSCAQESGMDRHPAACASARINV"
     CDS             complement(946..993)
                     /codon_start=1
                     /product="non-ORF yeast peptide conferring 
                     ubiquitin-dependent degradation"
                     /label=hCL1
                     /note="human codon-optimized"
                     /translation="ACKNWFSSLSHFVIHL"
     CDS             complement(1000..2646)
                     /codon_start=1
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="synthetic luc2 version of the luciferase gene"
                     /translation="EDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDAH
                     IEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAPA
                     NDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQSM
                     YTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHAR
                     DPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYKI
                     QSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGYG
                     LTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMSG
                     YVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESILL
                     QHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVVF
                     VDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     CDS             complement(2656..2676)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             complement(2701..2745)
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             complement(2767..2832)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     misc_feature    2833..3558
                     /label=per2 intron2
     CDS             complement(3559..3582)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        complement(3629..3647)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     promoter        complement(3653..3671)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     intron          3716..3848
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     protein_bind    complement(3913..3946)
                     /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(3997..4030)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     misc_feature    4055..4643
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(4526..4537)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4675..5151
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     repeat_region   5191..5331
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     rep_origin      5406..5861
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        6143..6247
                     /gene="bla"
                     /label=AmpR promoter
     CDS             6248..7108
                     /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      7279..7867
                     /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 cgcgtcggat taccgaggct ggtcacgtcg
      181 tcgcaggtga taggccggtg gccctggtct ctgccggctg tgagttgcgc agcggccaag
      241 caccattccc ccgcgccgca gtggtacgcg ccactccggg gctgcacgag cgggccaccg
      301 ccgtgccagg tgaatggaag tcccgcaggc cggaagtgga cgagcctact cgcccgggcg
      361 cgggggggcg caagagcgcg cagcatcttc attgaggaac ccgggcggcg aacatggagt
      421 tccatgtgcg tcttatgtaa agagagcgac gggcgtctcc accaattgat gagcgtagct
      481 ctcaggttcc gccccgccag tatgcaaatg aggtggcact ccgaccaatg gcgcgcgcag
      541 gggcgggctc agcgcgcgcg gtcacgtttt ccactatgtg acagcggagg gcgacgcggc
      601 ggcagcggcg ctactgggac tagcggctcc gggcggctgc ggcgcaggcc gagcgcacca
      661 agtgacgggc cgagcaaggg atctagagtc gactccggaa taacttcgta taggatactt
      721 tatacgaagt tatgcagaat ggtagctgga ttgtagctgc tattagcaat atgaaacctc
      781 ttaataactt cgtatagcat acattatacg aagttatggt tagacgttga tcctggcgct
      841 ggcgcaagca gcagggtgtc tatccatgcc gctctcctgg gcgcagctca tgggcagggt
      901 gccggcggcc tgctcctcca cctcgggagg gaagccgtgg ctgttaaggt ggatcacaaa
      961 gtggcttaag ctactgaacc agttcttgca agcagaattc acggcgatct tgccgccctt
     1021 cttggcctta atgagaatct cgcggatctt gcgggcgtcc aacttgccgg tcagtccttt
     1081 aggcacctcg tccacgaaca caacaccacc gcgcagcttc ttggcggttg taacctggct
     1141 ggccacatag tccacgatct ccttctcggt catggtttta ccgtgttcca gcacgacgac
     1201 tgcggcgggc agctcgccgg catcgtcgtc gggcaggccg gcgaccccgg cgtcgaagat
     1261 gttggggtgt tgcagcagga tgctctccag ttcggctggg gctacctggt agcccttgta
     1321 tttgatcagg ctcttcagcc ggtccacgat gaagaagtgc tcgtcctcgt cccagtaggc
     1381 gatgtcgccg ctgtgcagcc agccgtcctt gtcgatgaga gcgtttgtag cctcggggtt
     1441 gttaacgtag ccgctcatga tcatggggcc acggacgcac agctcgccgc gctggttcac
     1501 acccagtgtc ttaccggtgt ccaagtccac caccttagcc tcgaagaagg gcaccacctt
     1561 gcctactgcg ccaggcttgt cgtccccttc gggggtgatc agaatggcgc tggttgtttc
     1621 tgtcaggccg tagccctggc ggatgcctgg taggtggaag cgtttggcca cggcctcacc
     1681 tacctccttg ctgagcggcg ccccgccgct ggcgatctcg tgcaagttgc ttaggtcgta
     1741 cttgtcgatg agagtgctct tagcgaagaa gctaaatagt gtgggcacca gcagggcaga
     1801 ttgaatctta tagtcttgca agctgcgcaa gaatagctcc tcctcgaagc ggtacatgag
     1861 cacgacccga aagccgcaga tcaagtagcc cagcgtggtg aacatgccga agccgtggtg
     1921 aaatggcacc acgctgagga tagcggtgtc ggggatgatc tggttgccga agatggggtc
     1981 gcgggcatga ctgaatcgga cacaagcggt gcggtgcggt agggctacgc ccttgggcaa
     2041 tccggtactg ccactactgt tcatgatcag ggcgatggtt ttgtcccggt cgaagctctc
     2101 gggcacgaag tcgtactcgt tgaagccggg tggcaaatgg gaagtcacga aggtgtacat
     2161 gctttggaag ccctggtagt cggtcttgct atccatgatg atgatctttt gtatgatcgg
     2221 tagcttcttt tgcacgttga ggatcttttg cagccctttc ttgctcacga atacgacggt
     2281 gggctggctg atgcccatgc tgttcagcag ctcgcgctcg ttgtagatgt cgttagctgg
     2341 ggccacagcc acaccgatga acagggcacc caacacgggc atgaagaact gcaagctatt
     2401 ctcgctgcac accacgatcc gatggtttgt attcagccca tagcgcttca tagcttctgc
     2461 cagccgaacg ctcatctcga agtactcggc gtaggtaatg tccacctcga tatgtgcgtc
     2521 ggtaaaggcg atggtgccgg gcaccagggc gtagcgcttc atggctttgt gcagctgctc
     2581 gccggcggtc ccgtcttcga gtgggtagaa tggcgctggg cccttcttaa tgtttttggc
     2641 atcttcacct gcagggccct gaaaatacag gttttcggat cctgaaccgc ctgccgatcc
     2701 ctcatgccac tctattttct gagcttcgaa aatatcgttc aggccgctag ctgcggccgc
     2761 tccaccctta tcgtcatcgt ccttgtagtc gatgtcatga tctttataat caccgtcatg
     2821 gtctttgtag tcctgcaagg gaagatagat agataagatc tccacacgcc tcttccccca
     2881 accagccgat aaaactgacc agggacatca caccagagat tttaaagaaa atattaccgt
     2941 atgctgggtg gtggctttgg gacaaaggac ttgcctggag tgcctgagac ccatagtatg
     3001 gcctccatac ttcccctcaa aataaaagtc agagctggag aatcagctca atggtgaaga
     3061 gaagcaccca cacggctgct tacacccatc tgtaactcac agttccaggg gatccaatgc
     3121 cctcttctgt cccctatggt caccaaacat gcaagtggtg caccgacata catacaggca
     3181 aaacacccat atactttaaa gtaaaaattt aaaaaaggaa aagtcatgtt aaaactgaaa
     3241 cataagagct tcagctatta ggagacttga acactgagag acagaaatga aaaggaatat
     3301 tctgtgatcg aatgcatgtg tgtgagcagg ctcctccgag gtcttactgg actgtctgtt
     3361 aagtgcatca gatacaaaaa tgcaaagatg acgtggtatc gttatttgga aggacaaagt
     3421 acaataaata ccatctacag gattaagaag tactgccatc ctgcaagtcc tggtacttgc
     3481 tgcttgaaga cgggcgacaa actatggagg gagccacccg tgtcctgtgt gcgagggcct
     3541 ggcctgaggc tgacctacct tgtcatcgtc atccttgtag tccatggtgg cctctccttc
     3601 gagtgcaaaa agaacaagta gcttgtattc tatagtgtca cctaaatgct agcctatagt
     3661 gagtcgtatt aagtactcta gccttaagag ctgtaattga actgggagtg gacacctgtg
     3721 gagagaaagg caaagtggat gtcagtaaga ccaataggtg cctatcagaa acgcaagagt
     3781 cttctctgtc tcgacaagcc cagtttctat tggtctcctt aaacctgtct tgtaaccttg
     3841 atacttacct gcccagtgcc tcacgaccaa ctttcgctac cttaggaccg ttatagttac
     3901 tgcagcgcta gcataacttc gtataaagta tcctatacga agttatttgc cttaacccag
     3961 aaattatcac tgttattctt tagaatggtg caaagaataa cttcgtataa tgtatgctat
     4021 acgaagttat gaattcgata tcaagcttat cgataatcaa cctctggatt acaaaatttg
     4081 tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc
     4141 tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta
     4201 taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt
     4261 ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca
     4321 gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc
     4381 ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt
     4441 gtcggggaaa tcatcgtcct ttccttggct gctcgcctat gttgccacct ggattctgcg
     4501 cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg
     4561 cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat
     4621 ctccctttgg gccgcctccc cgcatcgata ccgagcgctg ctcgagagat ctacgggtgg
     4681 catccctgtg acccctcccc agtgcctctc ctggccctgg aagttgccac tccagtgccc
     4741 accagccttg tcctaataaa attaagttgc atcattttgt ctgactaggt gtccttctat
     4801 aatattatgg ggtggagggg ggtggtatgg agcaaggggc aagttgggaa gacaacctgt
     4861 agggcctgcg gggtctattg ggaaccaagc tggagtgcag tggcacaatc ttggctcact
     4921 gcaatctccg cctcctgggt tcaagcgatt ctcctgcctc agcctcccga gttgttggga
     4981 ttccaggcat gcatgaccag gctcagctaa tttttgtttt tttggtagag acggggtttc
     5041 accatattgg ccaggctggt ctccaactcc taatctcagg tgatctaccc accttggcct
     5101 cccaaattgc tgggattaca ggcgtgaacc actgctccct tccctgtcct tctgattttg
     5161 taggtaacca cgtgcggacc gagcggccgc aggaacccct agtgatggag ttggccactc
     5221 cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg
     5281 gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag gggcgcctga
     5341 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatacgt caaagcaacc
     5401 atagtacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     5461 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     5521 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     5581 atttagtgct ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag
     5641 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     5701 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcgggct attcttttga
     5761 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     5821 atttaacgcg aattttaaca aaatattaac gtttacaatt ttatggtgca ctctcagtac
     5881 aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc
     5941 gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg
     6001 gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct
     6061 cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg
     6121 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     6181 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     6241 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     6301 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     6361 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     6421 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     6481 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     6541 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     6601 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     6661 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     6721 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     6781 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     6841 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     6901 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     6961 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     7021 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     7081 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     7141 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     7201 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     7261 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     7321 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     7381 tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc
     7441 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     7501 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     7561 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     7621 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     7681 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     7741 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     7801 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     7861 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     7921 tcacatgt
//
