LOCUS       62056_7010        7579 bp DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7579)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7579
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             941..967
                     /codon_start=1
                     /label=HA
                     /note="HA (human influenza hemagglutinin) epitope tag"
                     /translation="YPYDVPDYA"
     CDS             968..2737
                     /codon_start=1
                     /label=hM3D(Gq)
                     /note="A modified form of human Gq-coupled M3 muscarinic 
                     receptor, belonging to a class of chemogenetic receptors 
                     called designer receptors exclusively activated by designer
                     drugs (DREADD)"
                     /translation="MTLHNNSTTSPLFPNISSSWIHSPSDAGLPPGTVTHFGSYNVSRA
                     AGNFSSPDGTTDDPLGGHTVWQVVFIAFLTGILALVTIIGNILVIVSFKVNKQLKTVNN
                     YFLLSLACADLIIGVISMNLFTTYIIMNRWALGNLACDLWLAIDCVASNASVMNLLVIS
                     FDRYFSITRPLTYRAKRTTKRAGVMIGLAWVISFVLWAPAILFWQYFVGKRTVPPGECF
                     IQFLSEPTITFGTAIAGFYMPVTIMTILYWRIYKETEKRTKELAGLQASGTEAETENFV
                     HPTGSSRSCSSYELQQQSMKRSNRRKYGRCHFWFTTKSWKPSSEQMDQDHSSSDSWNNN
                     DAAASLENSASSDEEDIGSETRAIYSIVLKLPGHSTILNSTKLPSSDNLQVPEEELGMV
                     DLERKADKLQAQKSVDDGGSFPKSFSKLPIQLESAVDTAKTSDVNSSVGKSTATLPLSF
                     KEATLAKRFALKTRSQITKRKRMSLVKEKKAAQTLSAILLAFIITWTPYNIMVLVNTFC
                     DSCIPKTFWNLGYWLCYINSTVNPVCYALCNKTFRTTFKMLLLCQCDKKKRRKQQYQQR
                     QSVIFHKRAPEQAL"
     CDS             2768..3484
                     /codon_start=1
                     /label=EGFP
                     /note="enhanced GFP"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     polyA_signal    3538..3762
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     protein_bind    4045..4092
                     /label=FRT
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             4100..5119
                     /codon_start=1
                     /label=HygR
                     /note="aminoglycoside phosphotransferase from E. coli"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     polyA_signal    5252..5385
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5422..5438)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(5446..5462)
                     /label=lac operator
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(5470..5500)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5515..5536)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(5824..6412)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6586..7443)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(7444..7548)
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttggta ccgagctcgg atccaccatg tacccatacg atgttccaga
      961 ttacgctatg accttgcaca ataacagtac aacctcgcct ttgtttccaa acatcagctc
     1021 ctcctggata cacagcccct ccgatgcagg gctgcccccg ggaaccgtca ctcatttcgg
     1081 cagctacaat gtttctcgag cagctggcaa tttctcctct ccagacggta ccaccgatga
     1141 ccctctggga ggtcataccg tctggcaagt ggtcttcatc gctttcttaa cgggcatcct
     1201 ggccttggtg accatcatcg gcaacatcct ggtaattgtg tcatttaagg tcaacaagca
     1261 gctgaagacg gtcaacaact acttcctctt aagcctggcc tgtgccgatc tgattatcgg
     1321 ggtcatttca atgaatctgt ttacgaccta catcatcatg aatcgatggg ccttagggaa
     1381 cttggcctgt gacctctggc ttgccattga ctgcgtagcc agcaatgcct ctgttatgaa
     1441 tcttctggtc atcagctttg acagatactt ttccatcacg aggccgctca cgtaccgagc
     1501 caaacgaaca acaaagagag ccggtgtgat gatcggtctg gcttgggtca tctcctttgt
     1561 cctttgggct cctgccatct tgttctggca atactttgtt ggaaagagaa ctgtgcctcc
     1621 gggagagtgc ttcattcagt tcctcagtga gcccaccatt acttttggca cagccatcgc
     1681 tggtttttat atgcctgtca ccattatgac tattttatac tggaggatct ataaggaaac
     1741 tgaaaagcgt accaaagagc ttgctggcct gcaagcctct gggacagagg cagagacaga
     1801 aaactttgtc caccccacgg gcagttctcg aagctgcagc agttacgaac ttcaacagca
     1861 aagcatgaaa cgctccaaca ggaggaagta tggccgctgc cacttctggt tcacaaccaa
     1921 gagctggaaa cccagctccg agcagatgga ccaagaccac agcagcagtg acagttggaa
     1981 caacaatgat gctgctgcct ccctggagaa ctccgcctcc tccgacgagg aggacattgg
     2041 ctccgagacg agagccatct actccatcgt gctcaagctt ccgggtcaca gcaccatcct
     2101 caactccacc aagttaccct catcggacaa cctgcaggtg cctgaggagg agctggggat
     2161 ggtggacttg gagaggaaag ccgacaagct gcaggcccag aagagcgtgg acgatggagg
     2221 cagttttcca aaaagcttct ccaagcttcc catccagcta gagtcagccg tggacacagc
     2281 taagacttct gacgtcaact cctcagtggg taagagcacg gccactctac ctctgtcctt
     2341 caaggaagcc actctggcca agaggtttgc tctgaagacc agaagtcaga tcactaagcg
     2401 gaaaaggatg tccctggtca aggagaagaa agcggcccag accctcagtg cgatcttgct
     2461 tgccttcatc atcacttgga ccccatacaa catcatggtt ctggtgaaca ccttttgtga
     2521 cagctgcata cccaaaacct tttggaatct gggctactgg ctgtgctaca tcaacagcac
     2581 cgtgaacccc gtgtgctatg ctctgtgcaa caaaacattc agaaccactt tcaagatgct
     2641 gctgctgtgc cagtgtgaca aaaaaaagag gcgcaagcag cagtaccagc agagacagtc
     2701 ggtcattttt cacaagcgcg cacccgagca ggccttgggt ggaggcggtt caggtggagg
     2761 tggctctatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga
     2821 gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc
     2881 cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg
     2941 gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct accccgacca
     3001 catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac
     3061 catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga
     3121 caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct
     3181 ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca
     3241 gaagaacggc atcaaggtga acttcaagat ccgccacaac atcgaggacg gcagcgtgca
     3301 gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga
     3361 caaccactac ctgagcaccc agtccgccct gagcaaagac cccaacgaga agcgcgatca
     3421 catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta
     3481 caagtaagcg gccgctcgag tctagagggc ccgtttaaac ccgctgatca gcctcgactg
     3541 tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg
     3601 aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga
     3661 gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg
     3721 aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa
     3781 ccagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg
     3841 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
     3901 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     3961 gggggtccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     4021 ttagggtgat ggttcacgta cctagaagtt cctattccga agttcctatt ctctagaaag
     4081 tataggaact tccttggcca aaaagcctga actcaccgcg acgtctgtcg agaagtttct
     4141 gatcgaaaag ttcgacagcg tctccgacct gatgcagctc tcggagggcg aagaatctcg
     4201 tgctttcagc ttcgatgtag gagggcgtgg atatgtcctg cgggtaaata gctgcgccga
     4261 tggtttctac aaagatcgtt atgtttatcg gcactttgca tcggccgcgc tcccgattcc
     4321 ggaagtgctt gacattgggg aattcagcga gagcctgacc tattgcatct cccgccgtgc
     4381 acagggtgtc acgttgcaag acctgcctga aaccgaactg cccgctgttc tgcagccggt
     4441 cgcggaggcc atggatgcga tcgctgcggc cgatcttagc cagacgagcg ggttcggccc
     4501 attcggaccg caaggaatcg gtcaatacac tacatggcgt gatttcatat gcgcgattgc
     4561 tgatccccat gtgtatcact ggcaaactgt gatggacgac accgtcagtg cgtccgtcgc
     4621 gcaggctctc gatgagctga tgctttgggc cgaggactgc cccgaagtcc ggcacctcgt
     4681 gcacgcggat ttcggctcca acaatgtcct gacggacaat ggccgcataa cagcggtcat
     4741 tgactggagc gaggcgatgt tcggggattc ccaatacgag gtcgccaaca tcttcttctg
     4801 gaggccgtgg ttggcttgta tggagcagca gacgcgctac ttcgagcgga ggcatccgga
     4861 gcttgcagga tcgccgcggc tccgggcgta tatgctccgc attggtcttg accaactcta
     4921 tcagagcttg gttgacggca atttcgatga tgcagcttgg gcgcagggtc gatgcgacgc
     4981 aatcgtccga tccggagccg ggactgtcgg gcgtacacaa atcgcccgca gaagcgcggc
     5041 cgtctggacc gatggctgtg tagaagtact cgccgatagt ggaaaccgac gccccagcac
     5101 tcgtccgagg gcaaaggaat agcacgtact acgagatttc gattccaccg ccgccttcta
     5161 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
     5221 ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta
     5281 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     5341 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag
     5401 ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     5461 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     5521 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     5581 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     5641 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     5701 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     5761 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     5821 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     5881 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     5941 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     6001 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     6061 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     6121 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     6181 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     6241 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt
     6301 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     6361 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     6421 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     6481 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     6541 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     6601 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     6661 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     6721 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     6781 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     6841 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     6901 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     6961 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     7021 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     7081 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     7141 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     7201 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     7261 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     7321 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     7381 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     7441 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     7501 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     7561 aaaagtgcca cctgacgtc
//
