LOCUS       V024935                 7812 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V024935
VERSION     V024935
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        50..429
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        430..633
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        678..696
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             756..2492
                     /label="NR1D2(NM_005126)"
                     /note="NR1D2(NM_005126)"
                     /gene="NR1D2"
     CDS             2499..2564
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2588..3176
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3242..3290
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3492..3920
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3934..4263
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4330..5124
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /note="confers resistance to neomycin, kanamycin, and G418
                     (Geneticin®)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /transl_table=1
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
                     /label="NeoR/KanR"
     polyA_signal    5300..5421
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5470..5486)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5494..5510
                     /bound_moiety="lac repressor encoded by lacI"
                     /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-β-D-thiogalactopyranoside (IPTG)."
                     /label="lac operator"
     promoter        complement(5518..5548)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5563..5584
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5872..6460)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6631..7491)
                     /codon_start=1
                     /gene="bla"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /product="β-lactamase"
                     /transl_table=1
                     /translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEY
                     SPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR
                     WEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS
                     ALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA
                     SLIKHW*"
                     /label="AmpR"
     promoter        complement(7492..7596)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta
       61 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
      121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      421 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      601 cgtgtacggt gggaggtcta tataagcaga gctctctggc taactagaga acccactgct
      661 tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagcgttta
      721 aacttaagct tggtaccgag ctcgaattcg ccaccatgga ggtgaatgca ggaggtgtga
      781 ttgcctatat cagttcttcc agctcagcct caagccctgc ctcttgtcac agtgagggtt
      841 ctgagaatag tttccagtcc tcctcctctt ctgttccatc ttctccaaat agctctaatt
      901 ctgataccaa tggtaatccc aagaatggtg atctcgccaa tattgaaggc atcttgaaga
      961 atgatcgaat agattgttct atgaaaacaa gcaaatcgag tgcacctggg atgacaaaaa
     1021 gtcatagtgg tgtgacaaaa tttagtggca tggttctact gtgtaaagtc tgtggggatg
     1081 tggcgtcagg attccactat ggagttcatg cttgcgaagg ctgtaagggt ttctttcgga
     1141 gaagtattca acaaaacatc cagtacaaga agtgcctgaa gaatgaaaac tgttctataa
     1201 tgagaatgaa taggaacaga tgtcagcaat gtcgcttcaa aaagtgtctg tctgttggaa
     1261 tgtcaagaga tgctgttcgg tttggtcgta ttcctaagcg tgaaaaacag aggatgctaa
     1321 ttgaaatgca aagtgcaatg aagaccatga tgaacagcca gttcagtggt cacttgcaaa
     1381 atgacacatt agtagaacat catgaacaga cagccttgcc agcccaggaa cagctgcgac
     1441 ccaagcccca actggagcaa gaaaacatca aaagctcttc tcctccatct tctgattttg
     1501 caaaggaaga agtgattggc atggtgacca gagctcacaa ggataccttt atgtataatc
     1561 aagagcagca agaaaactca gctgagagca tgcagcccca gagaggagaa cggattccca
     1621 agaacatgga gcaatataat ttaaatcatg atcattgcgg caatgggctt agcagccatt
     1681 ttccctgtag tgagagccag cagcatctca atggacagtt caaagggagg aatataatgc
     1741 attacccaaa tggtcatgcc atttgtattg caaatggaca ttgtatgaac ttctccaatg
     1801 cttatactca aagagtatgt gatagagttc cgatagatgg attttctcag aatgagaaca
     1861 agaatagtta cctgtgcaac actggaggaa gaatgcatct ggtttgtcca atgagtaagt
     1921 ctccatatgt ggatcctcat aaatcaggac atgaaatctg ggaagaattt tcgatgagct
     1981 tcactccagc agtgaaagaa gtggtggaat ttgcaaagcg tattcctggg ttcagagatc
     2041 tctctcagca tgaccaggtc aaccttttaa aggctgggac ttttgaggtt ttaatggtac
     2101 ggttcgcatc attatttgat gcaaaggaac gtactgtcac ctttttaagt ggaaagaaat
     2161 atagtgtgga tgatttacac tcaatgggag caggggatct gctaaactct atgtttgaat
     2221 ttagtgagaa gctaaatgcc ctccaactta gtgatgaaga gatgagtttg tttacagctg
     2281 ttgtcctggt atctgcagat cgatctggaa tagaaaacgt caactctgtg gaggctttgc
     2341 aggaaactct cattcgtgca ctaaggacct taataatgaa aaaccatcca aatgaggcct
     2401 ctatttttac aaaactgctt ctaaagttgc cagatcttcg atctttaaac aacatgcact
     2461 ctgaggagct cttggccttt aaagttcacc ctggatccga ctacaaagac catgacggtg
     2521 attataaaga tcatgacatc gactacaagg atgacgatga caagtagtga gggcccgata
     2581 tctcgacaat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta
     2641 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc
     2701 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga
     2761 ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac
     2821 ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc
     2881 cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc
     2941 tcggctgttg ggcactgaca attccgtggt gttgtcgggg aagctgacgt cctttccatg
     3001 gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc
     3061 ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc
     3121 gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcctgg
     3181 aaacggggga ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg
     3241 acggcaataa aaagacagaa taaaacgcac gggtgttggg tcgtttgttc ataaacgcgg
     3301 ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata
     3361 cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct
     3421 cgcagccaac gtcggggcgg caggccctgc catagcagat ctgcgcagct ggggctctag
     3481 ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     3541 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     3601 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcggggca tccctttagg
     3661 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     3721 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     3781 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     3841 ttttgattta taagggattt tggggatttc ggcctattgg ttaaaaaatg agctgattta
     3901 acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc
     3961 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
     4021 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     4081 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     4141 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     4201 tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     4261 aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga
     4321 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
     4381 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
     4441 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
     4501 aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
     4561 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
     4621 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
     4681 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
     4741 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
     4801 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc
     4861 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
     4921 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
     4981 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
     5041 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
     5101 cgccttcttg acgagttctt ctgagcggga ctctggggtt cgcgaaatga ccgaccaagc
     5161 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg
     5221 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     5281 ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa
     5341 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     5401 caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc
     5461 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     5521 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     5581 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     5641 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     5701 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     5761 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     5821 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     5881 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     5941 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     6001 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     6061 ttctcaatgc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     6121 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     6181 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     6241 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     6301 ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     6361 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     6421 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     6481 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     6541 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     6601 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     6661 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     6721 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     6781 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     6841 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     6901 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     6961 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     7021 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     7081 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     7141 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     7201 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     7261 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     7321 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     7381 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     7441 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     7501 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     7561 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     7621 tgacgtcgac ggatcgggag atctcccgat cccctatggt cgactctcag tacaatctgc
     7681 tctgatgccg catagttaag ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag
     7741 tagtgcgcga gcaaaattta agctacaaca aggcaaggct tgaccgacaa ttgcatgaag
     7801 aatctgctta gg
//
