LOCUS       Exported                6318 bp ds-DNA     circular SYN 25-JUL-2016
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6318)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 4, 2018 from SnapGene Viewer 4.1.7
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6318
                     /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"
     protein_bind    298..568
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     promoter        601..638
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     protein_bind    777..810
                     /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    861..894
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     CDS             complement(930..1643)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     misc_feature    complement(1644..2573)
                     /label=ChR2(H134R)
     protein_bind    complement(2580..2613)
                     /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(2664..2697)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     misc_feature    2722..3310
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(3193..3204)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3351..3558
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3581..3721
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     rep_origin      3796..4251
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        4533..4637
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4638..5498
                     /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      5669..6257
                     /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 cgcgaaacaa ttctgcagga attcggggcc
      181 gcggaggctg gatcggtccc ggtgtcttct atggaggtca aaacagcgtg gatggcgtct
      241 ccaggcgatc tgacggttca ctaaacgagc tctgcttata taggtcgagt ttaccactcc
      301 ctatcagtga tagagaaaag tgaaagtcga gtttaccact ccctatcagt gatagagaaa
      361 agtgaaagtc gagtttacca ctccctatca gtgatagaga aaagtgaaag tcgagtttac
      421 cactccctat cagtgataga gaaaagtgaa agtcgagttt accactccct atcagtgata
      481 gagaaaagtg aaagtcgagt ttaccactcc ctatcagtga tagagaaaag tgaaagtcga
      541 gtttaccact ccctatcagt gatagagaaa agtgaaagtc gagctcggta cccgggtcga
      601 gtaggcgtgt acggtgggag gcctatataa gcagagctcg tttagtgaac cgtcagatcg
      661 cctggagacg ccatccacgc tgttttgacc tccatagaag acaccgggac cgatccagcc
      721 tccgcggccc cgaattcgag ctcggtaccc ggggatcctc tagagtcgac tccggaataa
      781 cttcgtatag gatactttat acgaagttat gcagaatggt agctggattg tagctgctat
      841 tagcaatatg aaacctctta ataacttcgt atagcataca ttatacgaag ttatggcgcg
      901 cctcattaca cctcgttctc gtagcagaac ttgtacagct cgtccatgcc gagagtgatc
      961 ccggcggcgg tcacgaactc cagcaggacc atgtgatcgc gcttctcgtt ggggtctttg
     1021 ctcagggcgg actggtagct caggtagtgg ttgtcgggca gcagcacggg gccgtcgccg
     1081 atgggggtgt tctgctggta gtggtcggcg agctgcacgc tgccgtcctc gatgttgtgg
     1141 cggatcttga agttcacctt gatgccgttc ttctgcttgt cggccatgat atagacgttg
     1201 tggctgttgt agttgtactc cagcttgtgc cccaggatgt tgccgtcctc cttgaagtcg
     1261 atgcccttca gctcgatgcg gttcaccagg gtgtcgccct cgaacttcac ctcggcgcgg
     1321 gtcttgtagt tgccgtcgtc cttgaagaag atggtgcgct cctggacgta gccttcgggc
     1381 atggcggact tgaagaagtc gtgctgcttc atgtggtcgg ggtagcgggc gaagcactgc
     1441 aggccgtagc cgaaggtggt cacgagggtg ggccagggca cgggcagctt gccggtggtg
     1501 cagatgaact tcagggtcag cttgccgtag gtggcatcgc cctcgccctc gccggacacg
     1561 ctgaacttgt ggccgtttac gtcgccgtcc agctcgacca ggatgggcac caccccggtg
     1621 aacagctcct cgcccttgct cactggcacg gctccggcct cggcttcgtc ttcgacgaga
     1681 gtctcgacct cgatctccgt tccgccgatg ttcagtttgg tggttttgcg gatatctccg
     1741 tgaatcaata tgtgctcgtg gatcaggacg cgcaggtagt gtcccaacaa cccccaacaa
     1801 tttttactca tcagatcaat aatcgtgtga cctacggtgg agccatagac gctcaggacg
     1861 ccaaaacctt cgggccccaa aatgaagaga attgggaaca taccccagct cacgaaaaac
     1921 agccatgcca tgccggtcac gacctggcgg caccgaccct ttggcacagt atgataaccc
     1981 tcgatatatg ctttggcggc gtgaaaaaat gtgttcgcgc catagcacaa tccaagacaa
     2041 aagaagatga ctttaacata gccggttgcc atggcgctgg tagcccccca cacgatagtc
     2101 ccgatgtctg agacaaggag tcccatggtt ctcctgctgt agtcgttgct caggccggtg
     2161 aggttgctca ggtggataag gatgacagga caagtgagca gccactctgc atagcgcagc
     2221 cactgcaccc ggtgtcctgt ggcaaggtag agcatagagg gattcttaaa ctcaaaaaag
     2281 aactcgagaa tcaccttaac catttcaatg gcgcacacat agatctcctc ccagccgcat
     2341 gtagatttcc aggtttggta ggcatagaac atcagcagca aaatgctgaa tcctgctgca
     2401 agccactgca ggacatttga cgcggtctga gcgccgttcg tgccgcgaga ttcaatccat
     2461 ccggcacagt aacattgatc ctcagggacc aggacggacc cgttcaccac cacaggatta
     2521 gtaacgaaca aaagttcgcg tccgacggca gacaaagcgc cgccatagtc catgctagca
     2581 taacttcgta taaagtatcc tatacgaagt tatttgcctt aacccagaaa ttatcactgt
     2641 tattctttag aatggtgcaa agaataactt cgtataatgt atgctatacg aagttatgaa
     2701 ttcgatatca agcttatcga taatcaacct ctggattaca aaatttgtga aagattgact
     2761 ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg
     2821 tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg
     2881 ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg
     2941 tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct cctttccggg
     3001 actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc
     3061 tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaaatca
     3121 tcgtcctttc cttggctgct cgcctatgtt gccacctgga ttctgcgcgg gacgtccttc
     3181 tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct
     3241 ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc
     3301 gcctccccgc atcgataccg agcgctgctc gagagatcga tctgcctcga ctgtgccttc
     3361 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc
     3421 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg
     3481 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa
     3541 tagcaggcat gctggggaca cgtgcggacc gagcggccgc aggaacccct agtgatggag
     3601 ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc
     3661 cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag
     3721 gggcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatacgt
     3781 caaagcaacc atagtacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     3841 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     3901 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     3961 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg
     4021 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     4081 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcgggct
     4141 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
     4201 tttaacaaaa atttaacgcg aattttaaca aaatattaac gtttacaatt ttatggtgca
     4261 ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac
     4321 ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga
     4381 ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac
     4441 gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt
     4501 agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     4561 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     4621 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     4681 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     4741 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     4801 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     4861 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     4921 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     4981 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     5041 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     5101 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     5161 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     5221 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     5281 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     5341 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     5401 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     5461 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     5521 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     5581 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     5641 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     5701 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     5761 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc
     5821 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     5881 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     5941 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     6001 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     6061 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     6121 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     6181 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     6241 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     6301 ggccttttgc tcacatgt
//
