LOCUS       Exported                7688 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 7688)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Mar 27, 2018 from SnapGene 4.1.7
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7688
                     /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    150..1720
                     /label=P-CRY1
     misc_feature    663..667
                     /label=half CRE
     misc_feature    1296..1301
                     /label=e'box
     misc_feature    1315..1320
                     /label=E-box
     misc_feature    1367..1371
                     /label=half CRE
     misc_feature    1586..1590
                     /label=half CRE
     protein_bind    1733..1766
                     /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    1817..1850
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     misc_feature    1908..1915
                     /label=5'splice site
     intron          1910..2042
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     promoter        2087..2105
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        2111..2129
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     misc_feature    2168..2175
                     /label=KozaK
     CDS             2176..2199
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     misc_feature    2247..2582
                     /label=CRY1 intron336
     misc_feature    2407..2417
                     /label=RRE
     misc_feature    2462..2472
                     /label=RRE
     CDS             2590..2655
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             2677..2721
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             2746..2766
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             2776..3489
                     /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             3493..3513
                     /codon_start=1
                     /product="nuclear localization signal of SV40 large T 
                     antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     misc_feature    3532..3660
                     /label=D2
                     /note="destabilized by residues 422-461 of mouse ornithine 
                     decarboxylase, giving an in vivo half-life of ~2 hours"
     protein_bind    complement(3673..3706)
                     /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(3757..3790)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     misc_feature    3815..4403
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(4286..4297)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4435..4911
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     repeat_region   4951..5091
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     rep_origin      5166..5621
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        5903..6007
                     /gene="bla"
                     /label=AmpR promoter
     CDS             6008..6868
                     /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      7039..7627
                     /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 cgcgtgtaaa gatgcacatg tggccctgtg
      181 ttctcattag acttcaattg ctttgataaa acatcatgag caaaagcaac ttggccagga
      241 aggaaggggt ttgttgccgt ttacagttaa taatccatcg tggggagaag tcagggcaga
      301 aattcaaagc agaaacctgg agatagaggg ctgaagcaga agccatggag gaaagccgtt
      361 ttggttccct gtggcttcct cagcctgctt ccatatacaa cccgcctagg gctaccagcc
      421 taatggtggc gccacacaca ctggtcgggt ccttccacat cactcattaa tcaagaaagt
      481 gcccagactt ccctacaggc aacctgatgg aagtctttac tacaattaag gttcctcttc
      541 ccatttaact tcagttgaag aaaaaaaaaa aaaaaaaaaa aaaaagcaaa gcaaaaagcc
      601 caaaaaccta atcaaaatac cttctaaact acacttcatg taatagatta gagttgataa
      661 attgacgatg tatctacagt attatgaaat gctgaatatc tgagatcgga cctgtcttct
      721 ccagagcttg ggcaaaggtg cccgccccaa acagtaaaaa aggctaacgg cgtgatttct
      781 gctcctttca gagcagggga ctccgattaa cacctgattg aagctgggtg agaggcagga
      841 ctgtgcctcg gttgcctctc acccagccca gcaataactc tcagggcaca cggaactggc
      901 ttcagacctt ccacacaacc cataggaaga gccctgggcg cgctcccgcg tcctagcctc
      961 ttctcttcca ccgcgagagc gcagcgcacg cgcgccgtgc ccggcagagc gcgcgcccgc
     1021 gagctccccg cagccccgtg ctcatccggg tccccgcgag cctcggccaa tggaggcggc
     1081 tcccggcccc gggagcagaa ctatgcctcc tcccccttct cgcggcctgt ggcccccgcg
     1141 agcggcctgg ggctcaggag acccgcagag gctgcgagtc cggatcccac gcgagaactc
     1201 aggtcgtgag cgcccagcga tcgcaaccgg tcccgaagca ccgggcgcgg cggcgcacgc
     1261 gggggtctga gccagtgtag taaacacact tcagaaacgt gaggtgccgg tggtcacgtg
     1321 gggtgcatgc cgcccaatga gaagccgggg gcggggcgga ggccgctgac gcggcggagt
     1381 cacccgggca gcctcgggac cggtgaccgg caggcgagcg tcccgcggcc tggagctcta
     1441 gcccgcgctc tccgagctcg ctgcccactc cgggacgcac aggccgcgtc gcgcggaggc
     1501 cgcgtgagct cgtgtccgtt cgtggaggcc gcctcgggcg cgccaccgtc tcctcgcggg
     1561 tcgcctcggt tgggcgaggt ttccgtgacg agcctcccgg ggctgctgcg accagcccgg
     1621 gccgtcgtgg cggctcgggc cccgcgcgct cgcttgcgct ccgggggtct caggccctgg
     1681 aaggtatgcg tgtcctccgg acgagatctg cgatctaagt gtcgactccg gaataacttc
     1741 gtataggata ctttatacga agttatgcag aatggtagct ggattgtagc tgctattagc
     1801 aatatgaaac ctcttaataa cttcgtatag catacattat acgaagttat ggctgcagta
     1861 actataacgg tcctaaggta gcgaaagttg gtcgtgaggc actgggcagg taagtatcaa
     1921 ggttacaaga caggtttaag gagaccaata gaaactgggc ttgtcgagac agagaagact
     1981 cttgcgtttc tgataggcac ctattggtct tactgacatc cactttgcct ttctctccac
     2041 aggtgtccac tcccagttca attacagctc ttaaggctag agtacttaat acgactcact
     2101 ataggctagc atttaggtga cactatagaa tacaagctac ttgttctttt tgcactcgaa
     2161 ggagaggcca ccatggacta caaggatgac gatgacaaga attaccctgt tatccctaat
     2221 cgagatggtt gttatgacac agtgtagaaa ctatggcata ggacagatga ctgtgttata
     2281 tgaattcata atgtcttgtt ttcagtagca gtgggattat gttgtatctt gccattgcct
     2341 ttttcagact attttataga cagaactaga aatactagtt atataagctg tcctagcaca
     2401 gactagaaag taggtcattg tgatgggagt atgctaaacc acccactggt tgctatagcg
     2461 atgacctact ttagaaacta tttctgttga ttcatgttga aatcatcttt tggaagtggc
     2521 ataaggaagt tactacatgt acgtttttat agtagtaaaa ggtagagctg tcaagaggta
     2581 cccaccatgg actacaaaga ccatgacggt gattataaag atcatgacat cgactacaag
     2641 gacgatgacg ataagggtgg agcggccgca gctagcggcc tgaacgatat tttcgaagct
     2701 cagaaaatag agtggcatga gggatcggca ggcggttcag gatccgaaaa cctgtatttt
     2761 cagggccctg caggtgtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
     2821 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
     2881 gatgccacct acggcaagct gaccctgaag ttgatctgca ccaccggcaa gctgcccgtg
     2941 ccctggccca ccctcgtgac caccctcggc tacggcctgc agtgcttcgc ccgctacccc
     3001 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
     3061 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
     3121 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     3181 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat caccgccgac
     3241 aagcagaaga acggcatcaa ggcaaacttc aagatccgcc acaacatcga ggacggcggc
     3301 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     3361 cccgacaacc actacctgag ctaccagtcc aaactgagca aagaccccaa cgagaagcgc
     3421 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     3481 ctgtacaagc ctccaaaaaa gaagagaaag gtagaagacc ccgtgaattc taagcttagc
     3541 catggcttcc cgccggaggt ggaggagcag gatgatggca cgctgcccat gtcttgtgcc
     3601 caggagagcg ggatggaccg tcaccctgca gcctgtgctt ctgctaggat caatgtgtag
     3661 ggatccgcta gcataacttc gtataaagta tcctatacga agttatttgc cttaacccag
     3721 aaattatcac tgttattctt tagaatggtg caaagaataa cttcgtataa tgtatgctat
     3781 acgaagttat gaattcgata tcaagcttat cgataatcaa cctctggatt acaaaatttg
     3841 tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc
     3901 tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta
     3961 taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt
     4021 ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca
     4081 gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc
     4141 ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt
     4201 gtcggggaaa tcatcgtcct ttccttggct gctcgcctat gttgccacct ggattctgcg
     4261 cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg
     4321 cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat
     4381 ctccctttgg gccgcctccc cgcatcgata ccgagcgctg ctcgagagat ctacgggtgg
     4441 catccctgtg acccctcccc agtgcctctc ctggccctgg aagttgccac tccagtgccc
     4501 accagccttg tcctaataaa attaagttgc atcattttgt ctgactaggt gtccttctat
     4561 aatattatgg ggtggagggg ggtggtatgg agcaaggggc aagttgggaa gacaacctgt
     4621 agggcctgcg gggtctattg ggaaccaagc tggagtgcag tggcacaatc ttggctcact
     4681 gcaatctccg cctcctgggt tcaagcgatt ctcctgcctc agcctcccga gttgttggga
     4741 ttccaggcat gcatgaccag gctcagctaa tttttgtttt tttggtagag acggggtttc
     4801 accatattgg ccaggctggt ctccaactcc taatctcagg tgatctaccc accttggcct
     4861 cccaaattgc tgggattaca ggcgtgaacc actgctccct tccctgtcct tctgattttg
     4921 taggtaacca cgtgcggacc gagcggccgc aggaacccct agtgatggag ttggccactc
     4981 cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg
     5041 gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag gggcgcctga
     5101 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatacgt caaagcaacc
     5161 atagtacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     5221 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     5281 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     5341 atttagtgct ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag
     5401 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     5461 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcgggct attcttttga
     5521 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     5581 atttaacgcg aattttaaca aaatattaac gtttacaatt ttatggtgca ctctcagtac
     5641 aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc
     5701 gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg
     5761 gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct
     5821 cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg
     5881 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     5941 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     6001 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     6061 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     6121 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     6181 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     6241 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     6301 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     6361 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     6421 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     6481 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     6541 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     6601 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     6661 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     6721 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     6781 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     6841 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     6901 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     6961 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     7021 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     7081 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     7141 tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc
     7201 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     7261 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     7321 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     7381 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     7441 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     7501 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     7561 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     7621 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     7681 tcacatgt
//
