LOCUS       V036325                 8285 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V036325
VERSION     V036325
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             983..1006
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     CDS             1007..2659
                     /label="Gpr107(NM_178760)"
                     /note="Gpr107(NM_178760)"
                     /gene="Gpr107"
     CDS             2759..2779
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /transl_table=1
                     /translation="ENLYFQS"
                     /label="TEV site"
     CDS             2807..3808
                     /codon_start=1
                     /note="In the Tet-Off® system, tTA binds to the
                     Tet-responsive element&nbsp;and stimulates transcription
                     only in the absence of tetracycline or doxycycline."
                     /product="tetracycline-controlled transactivator,
                     comprising a fusion of the tetracycline repressor TetR with
                     the C-terminal activation domain of herpes simplex virus
                     VP16"
                     /transl_table=1
                     /translation="RLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHVKN
                     KRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTRPT
                     EKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTTDS
                     MPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS,AYSRARTKNNYGSTIE
                     GLLDLPDDDAPEEAGLAAPRLSFLPAGHTRRLSTAPPTDVSLGDELHLDGEDVAMAHAD
                     ALDDFDLDMLGDGDSPGPGFTPHDSAPYGALDMADFEFEQMFTDALGIDEYGG*"
                     /label="tTA"
     polyA_signal    3852..4076
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4122..4550
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4564..4893
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4960..5754
                     /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    5928..6049
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6098..6114)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6122..6138
                     /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(6146..6176)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6191..6212
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6500..7085)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7256..8116)
                     /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(8117..8221)
                     /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 cgccaccatg aagacgatca tcgccctgag ctacatcttc tgcctggtat tcgccgacta
      961 caaggacgat gatgacgcca gcgactacaa ggacgacgat gacaagatgg ccgtcccggt
     1021 acccctcggc cgcttcggct ccttctgcct gcggctgcta cgactgctgg cgttgctgga
     1081 gctgctggtc caccccgtct tgggccgcgt ccatcacctg gcgctcaaag atgatgtgcg
     1141 gcataaagtc catctcaaca ccttcgggtt cttcaaggat gggtacatgg tggtcaacgt
     1201 cagcagcctc tctgtgaacg agcctgaggg agccacggac aaggatgctg agatcggatt
     1261 cagtctggat cggactaaga acgatggctt ttcttcttac ctggatgaag atgtgaatta
     1321 ctgtatttta aaaaagaagt ccatgtcttc tgtcactctt gtcatcttag acatctctgg
     1381 aagcatagtc aaggtcagat ccccaccaga agctggtaag cagttaccag agattgtctt
     1441 cagcaaggat gagaaaatcc tgagtcagag ccaggagcct gctgtcagct ccaaccccaa
     1501 agacagtgag gctcggagaa ccctcgatgg tttcaaggct ggacgaagta cagtggactc
     1561 aaaggctatc acagagcgat ccttctcaat tcacaagaat gacggggtgg tctcatttca
     1621 gttcttcttt aacatcagca ccgatgacca ggaaggcctc tacagccttt acttccacaa
     1681 gtgctcaggc aacaacgtga agcctggtga acaggcctca ttcagcctga atattgctat
     1741 cacagagaag aaccctaaca gctacctctc tgccggggaa atccctctcc ccaagttata
     1801 tgtttccatg gccttgttct tctttctgtc tgggaccatt tggattcaca tccttcgtaa
     1861 gagaaggaat gatgtattta aaattcactg gttgatggcc gcccttcctt tcaccaagtc
     1921 tctctcgttg gtgttccatg caatcgacta ccactacatc tcctcgcagg gctttccgat
     1981 tgaaggctgg gctgttgtat actacataac tcacctgcta aagggcgcac tcctgttcat
     2041 caccatcgcc ctcatcggca ctggctgggc cttcatcaag cacatcctgt ctgataaaga
     2101 caagaagatc ttcatgattg tcatcccgct ccaggtcctg gcgaatgtgg cctacatcat
     2161 catagagtcc actgaggagg gcacaactga gtatggccta tggaaggact ctctgttcct
     2221 ggttgacttg ctttgctgtg gcgccatcct cttcccagtg gtgtggtcaa tcaggcattt
     2281 acaagaagcc tccgccacag atggaaaagc tgccattaac ttagcgaagc tgaggctttt
     2341 cagacattac tacgtcttga ttgtgtgcta catctacttt accaggatca tcgccttcct
     2401 tctcaagttt gctgttccat tccagtggaa gtggctctac cagctgctgg atgaaacagc
     2461 cacgctggtg ttctttgtcc tgacggggta taaatttcgc ccagcttcag ataaccccta
     2521 cctgcagctc tctcaggaag atgatgacct ggagatggag tccgtagtga cgacatcggg
     2581 tgtgatggag aatatgaaga aggtgaagaa agtgagcaac ggtgctgtgg agccccaggg
     2641 cagctgggaa ggcaccgcgg atatcatcga taccggtgga cgcaccccac ccagcctggg
     2701 tccccaagat gagtcctgca ccaccgccag ctcctccctg gccaaggaca cttcatcgga
     2761 aaacctgtac ttccaatcca ccggtgagaa cctgtacttc cagctaagat tagataaaag
     2821 taaagtgatt aacagcgcat tagagctgct taatgaggtc ggaatcgaag gtttaacaac
     2881 ccgtaaactc gcccagaagc taggtgtaga gcagcctaca ttgtattggc atgtaaaaaa
     2941 taagcgggct ttgctcgacg ccttagccat tgagatgtta gataggcacc atactcactt
     3001 ttgcccttta gaaggggaaa gctggcaaga ttttttacgt aataacgcta aaagttttag
     3061 atgtgcttta ctaagtcatc gcgatggagc aaaagtacat ttaggtacac ggcctacaga
     3121 aaaacagtat gaaactctcg aaaatcaatt agccttttta tgccaacaag gtttttcact
     3181 agagaatgca ttatatgcac tcagcgctgt ggggcatttt actttaggtt gcgtattgga
     3241 agatcaagag catcaagtcg ctaaagaaga aagggaaaca cctactactg atagtatgcc
     3301 gccattatta cgacaagcta tcgaattatt tgatcaccaa ggtgcagagc cagccttctt
     3361 attcggcctt gaattgatca tatgcggatt agaaaaacaa cttaaatgtg aaagtgggtc
     3421 cgcgtacagc cgcgcgcgta cgaaaaacaa ttacgggtct accatcgagg gcctgctcga
     3481 tctcccggac gacgacgccc ccgaagaggc ggggctggcg gctccgcgcc tgtcctttct
     3541 ccccgcggga cacacgcgca gactgtcgac ggcccccccg accgatgtca gcctggggga
     3601 cgagctccac ttagacggcg aggacgtggc gatggcgcat gccgacgcgc tagacgattt
     3661 cgatctggac atgttggggg acggggattc cccgggtccg ggatttaccc cccacgactc
     3721 cgccccctac ggcgctctgg atatggccga cttcgagttt gagcagatgt ttaccgatgc
     3781 ccttggaatt gacgagtacg gtgggtagct cgagtctaga gggcccgttt aaacccgctg
     3841 atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc
     3901 ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc
     3961 atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa
     4021 gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc
     4081 tgaggcggaa agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc
     4141 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     4201 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     4261 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     4321 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     4381 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     4441 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
     4501 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg
     4561 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     4621 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
     4681 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     4741 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     4801 ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga
     4861 ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt
     4921 ttcggatctg atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg
     4981 cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag
     5041 acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt
     5101 tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta
     5161 tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg
     5221 ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt
     5281 gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat
     5341 ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg
     5401 atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca
     5461 gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc
     5521 catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc
     5581 gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat
     5641 attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc
     5701 gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga
     5761 ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt
     5821 ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga
     5881 tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg
     5941 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     6001 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta
     6061 taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa
     6121 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     6181 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     6241 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     6301 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     6361 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     6421 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     6481 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     6541 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     6601 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     6661 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     6721 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     6781 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     6841 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     6901 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     6961 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     7021 ccaccgctgg tagcggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     7081 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     7141 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     7201 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     7261 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     7321 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     7381 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     7441 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     7501 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     7561 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     7621 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     7681 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     7741 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     7801 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     7861 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     7921 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     7981 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     8041 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     8101 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     8161 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     8221 gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc gggagatctc ccgatcccct
     8281 atggt
//
