LOCUS       Exported                6986 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 6986)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Mar 27, 2018 from SnapGene 4.1.7
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6986
                     /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)."
     misc_feature    complement(820..948)
                     /label=D2
                     /note="destabilized by residues 422-461 of mouse ornithine 
                     decarboxylase, giving an in vivo half-life of ~2 hours"
     CDS             complement(967..987)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 large T 
                     antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(991..1704)
                     /codon_start=1
                     /product="Venus YFP with monomerizing A206K mutation (Nagai
                     et al., 2002)"
                     /label=mVenus
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     LICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             complement(1714..1734)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             complement(1759..1803)
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             complement(1825..1890)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     misc_feature    1891..2616
                     /label=per2 intron2
     CDS             complement(2617..2640)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        complement(2687..2705)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     promoter        complement(2711..2729)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     intron          2774..2906
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     misc_feature    2901..2908
                     /label=5' splice site
     protein_bind    complement(2971..3004)
                     /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(3055..3088)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     misc_feature    3113..3701
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(3584..3595)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3733..4209
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     repeat_region   4249..4389
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     rep_origin      4464..4919
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        5201..5305
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5306..6166
                     /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      6337..6925
                     /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 aagttatggc tacacattga tcctagcaga
      841 agcacaggct gcagggtgac ggtccatccc gctctcctgg gcacaagaca tgggcagcgt
      901 gccatcatcc tgctcctcca cctccggcgg gaagccatgg ctaagcttag aattcacggg
      961 gtcttctacc tttctcttct tttttggagg cttgtacagc tcgtccatgc cgagagtgat
     1021 cccggcggcg gtcacgaact ccagcaggac catgtgatcg cgcttctcgt tggggtcttt
     1081 gctcagtttg gactggtagc tcaggtagtg gttgtcgggc agcagcacgg ggccgtcgcc
     1141 gatgggggtg ttctgctggt agtggtcggc gagctgcacg ccgccgtcct cgatgttgtg
     1201 gcggatcttg aagtttgcct tgatgccgtt cttctgcttg tcggcggtga tatagacgtt
     1261 gtggctgttg tagttgtact ccagcttgtg ccccaggatg ttgccgtcct ccttgaagtc
     1321 gatgcccttc agctcgatgc ggttcaccag ggtgtcgccc tcgaacttca cctcggcgcg
     1381 ggtcttgtag ttgccgtcgt ccttgaagaa gatggtgcgc tcctggacgt agccttcggg
     1441 catggcggac ttgaagaagt cgtgctgctt catgtggtcg gggtagcggg cgaagcactg
     1501 caggccgtag ccgagggtgg tcacgagggt gggccagggc acgggcagct tgccggtggt
     1561 gcagatcaac ttcagggtca gcttgccgta ggtggcatcg ccctcgccct cgccggacac
     1621 gctgaacttg tggccgttta cgtcgccgtc cagctcgacc aggatgggca ccaccccggt
     1681 gaacagctcc tcgcccttgc tcacacctgc agggccctga aaatacaggt tttcggatcc
     1741 tgaaccgcct gccgatccct catgccactc tattttctga gcttcgaaaa tatcgttcag
     1801 gccgctagct gcggccgctc cacccttatc gtcatcgtcc ttgtagtcga tgtcatgatc
     1861 tttataatca ccgtcatggt ctttgtagtc ctgcaaggga agatagatag ataagatctc
     1921 cacacgcctc ttcccccaac cagccgataa aactgaccag ggacatcaca ccagagattt
     1981 taaagaaaat attaccgtat gctgggtggt ggctttggga caaaggactt gcctggagtg
     2041 cctgagaccc atagtatggc ctccatactt cccctcaaaa taaaagtcag agctggagaa
     2101 tcagctcaat ggtgaagaga agcacccaca cggctgctta cacccatctg taactcacag
     2161 ttccagggga tccaatgccc tcttctgtcc cctatggtca ccaaacatgc aagtggtgca
     2221 ccgacataca tacaggcaaa acacccatat actttaaagt aaaaatttaa aaaaggaaaa
     2281 gtcatgttaa aactgaaaca taagagcttc agctattagg agacttgaac actgagagac
     2341 agaaatgaaa aggaatattc tgtgatcgaa tgcatgtgtg tgagcaggct cctccgaggt
     2401 cttactggac tgtctgttaa gtgcatcaga tacaaaaatg caaagatgac gtggtatcgt
     2461 tatttggaag gacaaagtac aataaatacc atctacagga ttaagaagta ctgccatcct
     2521 gcaagtcctg gtacttgctg cttgaagacg ggcgacaaac tatggaggga gccacccgtg
     2581 tcctgtgtgc gagggcctgg cctgaggctg acctaccttg tcatcgtcat ccttgtagtc
     2641 catggtggcc tctccttcga gtgcaaaaag aacaagtagc ttgtattcta tagtgtcacc
     2701 taaatgctag cctatagtga gtcgtattaa gtactctagc cttaagagct gtaattgaac
     2761 tgggagtgga cacctgtgga gagaaaggca aagtggatgt cagtaagacc aataggtgcc
     2821 tatcagaaac gcaagagtct tctctgtctc gacaagccca gtttctattg gtctccttaa
     2881 acctgtcttg taaccttgat acttacctgc ccagtgcctc acgaccaact ttcgctacct
     2941 taggaccgtt atagttactg cagcgctagc ataacttcgt ataaagtatc ctatacgaag
     3001 ttatttgcct taacccagaa attatcactg ttattcttta gaatggtgca aagaataact
     3061 tcgtataatg tatgctatac gaagttatga attcgatatc aagcttatcg ataatcaacc
     3121 tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac
     3181 gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt
     3241 cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt
     3301 tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg
     3361 cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac
     3421 ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac
     3481 tgacaattcc gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctatgt
     3541 tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc
     3601 ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg
     3661 ccctcagacg agtcggatct ccctttgggc cgcctccccg catcgatacc gagcgctgct
     3721 cgagagatct acgggtggca tccctgtgac ccctccccag tgcctctcct ggccctggaa
     3781 gttgccactc cagtgcccac cagccttgtc ctaataaaat taagttgcat cattttgtct
     3841 gactaggtgt ccttctataa tattatgggg tggagggggg tggtatggag caaggggcaa
     3901 gttgggaaga caacctgtag ggcctgcggg gtctattggg aaccaagctg gagtgcagtg
     3961 gcacaatctt ggctcactgc aatctccgcc tcctgggttc aagcgattct cctgcctcag
     4021 cctcccgagt tgttgggatt ccaggcatgc atgaccaggc tcagctaatt tttgtttttt
     4081 tggtagagac ggggtttcac catattggcc aggctggtct ccaactccta atctcaggtg
     4141 atctacccac cttggcctcc caaattgctg ggattacagg cgtgaaccac tgctcccttc
     4201 cctgtccttc tgattttgta ggtaaccacg tgcggaccga gcggccgcag gaacccctag
     4261 tgatggagtt ggccactccc tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa
     4321 aggtcgcccg acgcccgggc tttgcccggg cggcctcagt gagcgagcga gcgcgcagct
     4381 gcctgcaggg gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc
     4441 gcatacgtca aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     4501 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     4561 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     4621 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgattt
     4681 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     4741 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     4801 ctcgggctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
     4861 tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt
     4921 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc
     4981 gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca
     5041 agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg
     5101 cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat
     5161 ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     5221 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     5281 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     5341 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     5401 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     5461 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     5521 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     5581 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     5641 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     5701 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     5761 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     5821 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     5881 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     5941 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     6001 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     6061 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     6121 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     6181 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     6241 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     6301 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     6361 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     6421 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     6481 tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     6541 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     6601 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     6661 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     6721 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     6781 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     6841 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     6901 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     6961 cttttgctgg ccttttgctc acatgt
//
