LOCUS       Exported                7683 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 7683)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Mar 27, 2018 from SnapGene 4.1.7
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7683
                     /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
     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    complement(1853..1981)
                     /label=D2
                     /note="destabilized by residues 422-461 of mouse ornithine 
                     decarboxylase, giving an in vivo half-life of ~2 hours"
     CDS             complement(2000..2020)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 large T 
                     antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(2024..2737)
                     /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(2747..2767)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             complement(2792..2836)
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             complement(2858..2923)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     misc_feature    2931..3266
                     /label=Cry1 intron336
     CDS             complement(3314..3337)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        complement(3384..3402)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     promoter        complement(3408..3426)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     intron          3471..3603
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     misc_feature    complement(3598..3605)
                     /label=5' splice site
     protein_bind    complement(3668..3701)
                     /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(3752..3785)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     misc_feature    3810..4398
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(4281..4292)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4430..4906
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     repeat_region   4946..5086
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     rep_origin      5161..5616
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        5898..6002
                     /gene="bla"
                     /label=AmpR promoter
     CDS             6003..6863
                     /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      7034..7622
                     /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 ggctacacat
     1861 tgatcctagc agaagcacag gctgcagggt gacggtccat cccgctctcc tgggcacaag
     1921 acatgggcag cgtgccatca tcctgctcct ccacctccgg cgggaagcca tggctaagct
     1981 tagaattcac ggggtcttct acctttctct tcttttttgg aggcttgtac agctcgtcca
     2041 tgccgagagt gatcccggcg gcggtcacga actccagcag gaccatgtga tcgcgcttct
     2101 cgttggggtc tttgctcagt ttggactggt agctcaggta gtggttgtcg ggcagcagca
     2161 cggggccgtc gccgatgggg gtgttctgct ggtagtggtc ggcgagctgc acgccgccgt
     2221 cctcgatgtt gtggcggatc ttgaagtttg ccttgatgcc gttcttctgc ttgtcggcgg
     2281 tgatatagac gttgtggctg ttgtagttgt actccagctt gtgccccagg atgttgccgt
     2341 cctccttgaa gtcgatgccc ttcagctcga tgcggttcac cagggtgtcg ccctcgaact
     2401 tcacctcggc gcgggtcttg tagttgccgt cgtccttgaa gaagatggtg cgctcctgga
     2461 cgtagccttc gggcatggcg gacttgaaga agtcgtgctg cttcatgtgg tcggggtagc
     2521 gggcgaagca ctgcaggccg tagccgaggg tggtcacgag ggtgggccag ggcacgggca
     2581 gcttgccggt ggtgcagatc aacttcaggg tcagcttgcc gtaggtggca tcgccctcgc
     2641 cctcgccgga cacgctgaac ttgtggccgt ttacgtcgcc gtccagctcg accaggatgg
     2701 gcaccacccc ggtgaacagc tcctcgccct tgctcacacc tgcagggccc tgaaaataca
     2761 ggttttcgga tcctgaaccg cctgccgatc cctcatgcca ctctattttc tgagcttcga
     2821 aaatatcgtt caggccgcta gctgcggccg ctccaccctt atcgtcatcg tccttgtagt
     2881 cgatgtcatg atctttataa tcaccgtcat ggtctttgta gtccatggtg ggtacctctt
     2941 gacagctcta ccttttacta ctataaaaac gtacatgtag taacttcctt atgccacttc
     3001 caaaagatga tttcaacatg aatcaacaga aatagtttct aaagtaggtc atcgctatag
     3061 caaccagtgg gtggtttagc atactcccat cacaatgacc tactttctag tctgtgctag
     3121 gacagcttat ataactagta tttctagttc tgtctataaa atagtctgaa aaaggcaatg
     3181 gcaagataca acataatccc actgctactg aaaacaagac attatgaatt catataacac
     3241 agtcatctgt cctatgccat agtttctaca ctgtgtcata acaaccatct cgattaggga
     3301 taacagggta attcttgtca tcgtcatcct tgtagtccat ggtggcctct ccttcgagtg
     3361 caaaaagaac aagtagcttg tattctatag tgtcacctaa atgctagcct atagtgagtc
     3421 gtattaagta ctctagcctt aagagctgta attgaactgg gagtggacac ctgtggagag
     3481 aaaggcaaag tggatgtcag taagaccaat aggtgcctat cagaaacgca agagtcttct
     3541 ctgtctcgac aagcccagtt tctattggtc tccttaaacc tgtcttgtaa ccttgatact
     3601 tacctgccca gtgcctcacg accaactttc gctaccttag gaccgttata gttactgcag
     3661 cgctagcata acttcgtata aagtatccta tacgaagtta tttgccttaa cccagaaatt
     3721 atcactgtta ttctttagaa tggtgcaaag aataacttcg tataatgtat gctatacgaa
     3781 gttatgaatt cgatatcaag cttatcgata atcaacctct ggattacaaa atttgtgaaa
     3841 gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa
     3901 tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat
     3961 cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt
     4021 gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc
     4081 tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc
     4141 ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg
     4201 ggaaatcatc gtcctttcct tggctgctcg cctatgttgc cacctggatt ctgcgcggga
     4261 cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc
     4321 tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc
     4381 tttgggccgc ctccccgcat cgataccgag cgctgctcga gagatctacg ggtggcatcc
     4441 ctgtgacccc tccccagtgc ctctcctggc cctggaagtt gccactccag tgcccaccag
     4501 ccttgtccta ataaaattaa gttgcatcat tttgtctgac taggtgtcct tctataatat
     4561 tatggggtgg aggggggtgg tatggagcaa ggggcaagtt gggaagacaa cctgtagggc
     4621 ctgcggggtc tattgggaac caagctggag tgcagtggca caatcttggc tcactgcaat
     4681 ctccgcctcc tgggttcaag cgattctcct gcctcagcct cccgagttgt tgggattcca
     4741 ggcatgcatg accaggctca gctaattttt gtttttttgg tagagacggg gtttcaccat
     4801 attggccagg ctggtctcca actcctaatc tcaggtgatc tacccacctt ggcctcccaa
     4861 attgctggga ttacaggcgt gaaccactgc tcccttccct gtccttctga ttttgtaggt
     4921 aaccacgtgc ggaccgagcg gccgcaggaa cccctagtga tggagttggc cactccctct
     4981 ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt
     5041 gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc tgcaggggcg cctgatgcgg
     5101 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag caaccatagt
     5161 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     5221 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     5281 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     5341 gtgctttacg gcacctcgac cccaaaaaac ttgatttggg tgatggttca cgtagtgggc
     5401 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     5461 gactcttgtt ccaaactgga acaacactca accctatctc gggctattct tttgatttat
     5521 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
     5581 acgcgaattt taacaaaata ttaacgttta caattttatg gtgcactctc agtacaatct
     5641 gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct
     5701 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct
     5761 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga
     5821 tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca
     5881 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     5941 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     6001 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     6061 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     6121 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     6181 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     6241 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     6301 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     6361 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     6421 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     6481 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     6541 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     6601 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     6661 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     6721 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     6781 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     6841 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     6901 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     6961 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     7021 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     7081 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     7141 aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt
     7201 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     7261 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     7321 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     7381 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     7441 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     7501 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     7561 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     7621 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     7681 tgt
//
