LOCUS       V034431                 7975 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V034431
VERSION     V034431
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        202..581
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        582..785
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        830..848
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             931..3468
                     /label="CLOCK(NM_001267843)"
                     /note="CLOCK(NM_001267843)"
                     /gene="CLOCK"
     CDS             3478..3495
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /transl_table=1
                     /translation="HHHHHH"
                     /label="6xHis"
     polyA_signal    3542..3766
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3812..4240
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4254..4583
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4650..5444
                     /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    5618..5739
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5788..5804)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5812..5828
                     /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(5836..5866)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5881..5902
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6190..6775)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6946..7806)
                     /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(7807..7911)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
       61 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
      121 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg
      181 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta
      241 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      301 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      361 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      421 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
      481 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
      541 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      601 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      661 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
      721 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      781 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc
      841 actataggga gacccaagct ggctagcgtt taaacttaag cttggtaccg agctcggatc
      901 cactagtcca gtgtggtgga attcgccacc atgttgttta ccgtaagctg tagtaaaatg
      961 agctcgattg ttgacagaga tgacagtagt atttttgatg ggttggtgga agaagatgac
     1021 aaggacaaag cgaaaagagt atctagaaac aaatctgaaa agaaacgtag agatcaattt
     1081 aatgttctca ttaaagaact gggatccatg cttcctggta atgctagaaa gatggacaaa
     1141 tctactgttc tgcagaaaag cattgatttt ttacgaaaac ataaagaaat cactgcacag
     1201 tcagatgcta gtgaaattcg acaggactgg aaacctacat tccttagtaa tgaagagttt
     1261 acacaattaa tgttagaggc tcttgatggt ttttttttag caatcatgac agatggaagc
     1321 ataatatatg tgtctgagag tgtaacttca ttacttgaac atttaccatc tgatcttgtg
     1381 gatcaaagta tatttaattt tatcccagaa ggggaacatt cagaggttta taaaatactc
     1441 tctactcatc tgctggaaag tgattcatta accccagaat atttaaaatc aaaaaatcag
     1501 ttagaattct gttgtcacat gctgcgagga acaatagacc caaaggagcc atctacctat
     1561 gaatatgtaa aatttatagg aaatttcaaa tctttaaaca gtgtatcctc ttcagcacac
     1621 aatggttttg aaggaactat acaacgcaca cataggccat cttatgaaga tagagtttgt
     1681 tttgtagcta ctgtcaggtt agctacacct cagttcatca aggaaatgtg cactgttgaa
     1741 gaacccaatg aagagtttac atctagacat agtttagaat ggaagtttct gtttctagat
     1801 cacagggcac cacccataat agggtatttg ccatttgaag ttctgggaac atcaggctat
     1861 gattactatc atgtggatga cctagaaaat ttggcaaaat gtcatgagca cttaatgcaa
     1921 tatgggaaag gcaaatcatg ttattatagg ttcctgacta aggggcaaca gtggatttgg
     1981 cttcagactc attattatat cacttaccat cagtggaatt caaggccaga gtttattgtt
     2041 tgtactcaca ctgtagtaag ttatgcagaa gttagggctg aaagacgacg agaacttggc
     2101 attgaagagt ctcttcctga gacagctgct gacaaaagcc aagattctgg gtcagataat
     2161 cgtataaaca cagtcagtct caaggaagca ttggaaaggt ttgatcacag cccaacccct
     2221 tctgcctctt ctcggagttc aagaaaatca tctcacacgg ccgtctcaga cccttcctca
     2281 acaccaacca agatcccgac ggatacgagc actccaccca ggcagcattt accagctcat
     2341 gagaagatgg tgcaaagaag gtcatcattt agtagtcagt ccataaattc ccagtctgtt
     2401 ggttcatcat taacacagcc agtgatgtct caagctacaa atttaccaat tccacaaggc
     2461 atgtcccagt ttcagttttc agctcaatta ggagccatgc aacatctgaa agaccaattg
     2521 gaacaacgga cacgcatgat agaagcaaat attcatcggc aacaagaaga actaagaaaa
     2581 attcaagaac aacttcagat ggtccatggt caggggctgc agatgttttt gcaacaatca
     2641 aatcctgggt tgaattttgg ttccgttcaa ctttcttctg gaaattcatc taatatccag
     2701 caacttgcac ctataaatat gcaaggccaa gttgttccta ctaaccagat tcaaagtgga
     2761 atgaatactg gacacattgg cacaactcag cacatgatac aacaacagac tttacagagt
     2821 acatcaactc agagtcaaca aaatgtactg agtgggcaca gtcagcaaac atctctaccc
     2881 agtcagacac agagcactct tacagcccca ctgtataaca ctatggtgat ttctcagcct
     2941 gcagccggaa gcatggtcca gattccatct agtatgccac aaaacagcac ccagagtgct
     3001 gcagtaacta cattcactca ggacaggcag ataagatttt ctcaaggtca acaacttgtg
     3061 accaaattag tgactgctcc tgtagcttgt ggggcagtca tggtacctag tactatgctt
     3121 atgggccagg tggtgactgc atatcctact tttgctacac aacagcaaca gtcacagaca
     3181 ttgtcagtaa cgcagcagca gcagcagcag agctcccagg agcagcagct cacttcagtt
     3241 cagcaaccat ctcaggctca gctgacccag ccaccgcaac aatttttaca gacttctagg
     3301 ttgctccatg ggaatccctc aactcaactc attctctctg ctgcatttcc tctacaacag
     3361 agcaccttcc ctcagtcaca tcaccagcaa catcagtctc agcaacagca gcaactcagc
     3421 cggcacagga ctgacagctt gcccgaccct tccaaggttc aaccacaggc ggccgcacat
     3481 caccatcatc accattgact cgagtctaga gggcccgttt aaacccgctg atcagcctcg
     3541 actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc
     3601 ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt
     3661 ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat
     3721 tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa
     3781 agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg
     3841 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     3901 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     3961 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     4021 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     4081 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     4141 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     4201 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc
     4261 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     4321 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     4381 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     4441 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     4501 atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt
     4561 ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg
     4621 atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt
     4681 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct
     4741 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga
     4801 ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg
     4861 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact
     4921 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg
     4981 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct
     5041 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg
     5101 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt
     5161 tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg
     5221 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc
     5281 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag
     5341 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt
     5401 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt
     5461 cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc
     5521 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
     5581 gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa
     5641 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     5701 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac
     5761 ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     5821 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     5881 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     5941 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     6001 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     6061 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     6121 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     6181 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     6241 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     6301 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     6361 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     6421 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     6481 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     6541 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     6601 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     6661 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     6721 tagcggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     6781 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     6841 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     6901 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     6961 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     7021 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     7081 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     7141 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     7201 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     7261 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     7321 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     7381 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     7441 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     7501 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     7561 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     7621 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     7681 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     7741 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     7801 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     7861 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     7921 ccgaaaagtg ccacctgacg tcgacggatc gggagatctc ccgatcccct atggt
//
