LOCUS       V034845                 8416 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V034845
VERSION     V034845
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        235..614
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        615..818
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        863..881
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             964..3906
                     /label="Nr3c2(NM_013131)"
                     /note="Nr3c2(NM_013131)"
                     /gene="Nr3c2"
     CDS             3913..3978
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    4016..4240
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4286..4714
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4728..5057
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5124..5918
                     /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    6092..6213
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6262..6278)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6286..6302
                     /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(6310..6340)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6355..6376
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6664..7249)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7420..8280)
                     /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(8281..8385)
                     /gene="bla"
                     /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 atccactagt ccagtgtggt ggaattcgcc
      961 accatggaaa ccaaaggcta ccacagtctc cctgaaggcc tagacatgga aaggcgctgg
     1021 agtcaagtgt ctcagacctt ggagcgttct tcacttggac ctgcagagag gaccactgag
     1081 aacaactaca tggagattgt caacgtcagc tgcgtttccg gtgctattcc gaacaacagt
     1141 actcaaggaa gcagcaaaga aaagcacgaa ttactccctt gcattcagca agacaatagt
     1201 cggtctggga ttttgccatc agatattaaa actgagctgg aatccaagga actttcagcc
     1261 acggtagctg agtccatggg tttatatatg gattctgtga gagatgccga gtacacttac
     1321 gatcagcaga accaacaagg aagcctgagc ccaacaaaga tttatcaaaa catggagcag
     1381 ctggtgaagt tttacaaaga gaatggtcac aggtcctcca cactgagtgc tatgagcaga
     1441 cctttgaggt cattcatgcc tgactctgca gcctccatga atggtggggc cttgcgtgcc
     1501 atcgtgaaga gccctatcat ctgtcatgag aagagctctt ctgttagcag cccgctgaac
     1561 atggcatctt cagtatgcag ccctgtgggc atcaactcca tgtcctcctc cacgactagc
     1621 tttggcagtt tcccagtgca cagtcccatc actcaaggaa cctcactgac atgctccccc
     1681 agtgtcgaaa acagaggctc aaggtcacac agccccacac atgcgagcaa cgtgggctcc
     1741 cctctttcaa gtccattgag cagcatgaaa tctccaatct ccagccctcc aagtcattgc
     1801 agtgtaaaat ctccagtctc cagtccaaat aacgtccctc tgcgctcctc tgtgtccagc
     1861 ccggcaaatc tcaacaactc aaggtgctcc gtttccagcc cttccaacaa caccaacaat
     1921 agatccacac tctccagccc gacagccagc acagtggggt ccatcggcag ccccatcagc
     1981 aatgccttca gctatgccac ttcaggcgct tctgctggag ccggtgccat ccaggatgtg
     2041 gttcctagtc cagacaccca cgagaaaggt gctcacgacg ttcctttccc taagacagag
     2101 gaagtcgaga aggccatctc caatggtgtg actggcccgc tcaacattgt ccagtacata
     2161 aaatcagaac cagatggggc tttcagcagt tcctgtctag gaggaaacag caaaatcagc
     2221 cccagttctc cattctctgt accaataaag caagagtcaa gcaagcactc atgttcaggc
     2281 gcctctttta aagggaaccc cacggtcaac ccatttccat tcatggatgg ctcctacttt
     2341 tcctttatgg atgataagga ctattattcc ctatcaggaa tcttaggacc acctgtgccc
     2401 ggctttgatg gtagctgcga gggcagcgca ttcccagtgg ggattaagca agaaccagat
     2461 gatgggagct attaccctga agccagcatc ccatcatctg ctatcgttgg tgtgaattcg
     2521 ggtggacagt cctttcacta ccggattggt gctcaaggta caatatcttt atcacggtca
     2581 cctagagacc aatctttcca acacttgagt tccttcccac ctgtcaatac attagtggaa
     2641 tcatggaaac cacacggtga cctgtcatct aggagaagtg atgggtatcc cgtcctagag
     2701 tacattccag aaaacgtgtc aagctctact ttacgaagtg tttctactgg atcctcaaga
     2761 ccttccaaga tctgtttggt gtgtggagat gaggcttctg ggtgtcacta tggggtagtg
     2821 acctgtggca gctgcaaagt cttcttcaaa agagccgtgg aagggcaaca caactatctg
     2881 tgtgctggaa gaaatgactg cattattgat aagattcggc gaaagaactg tcctgcctgc
     2941 aggctccaga aatgccttca agctggcatg aacttaggag ctcgaaagtc aaagaagctg
     3001 gggaagttaa aaggcctcca cgaggagcaa ccacagcagc ccccaccacc gccaccccag
     3061 agtccagaag aggggaccac atacatcgct ccgaccaagg agccatcggt gaactctgcg
     3121 ctggtcccgc agctcacctc cattacgcat gcactcacac catccccggc catgatcctg
     3181 gagaacatcg agcctgagac cgtgtatgca ggctacgaca attccaagcc cgacaccgcc
     3241 gagagcctgc tctccacgct caaccgcctg gcaggcaaac agatgatcca ggtcgtgaag
     3301 tgggccaagg tacttccagg atttaaaaac ttgcctctcg aggaccaaat caccctcatc
     3361 cagtattctt ggatgtgtct atcatcgttc gctttgagtt ggagatcgta caaacacacg
     3421 aacagccaac tcctctattt tgctccagac ctagttttta atgaagagaa gatgcatcag
     3481 tctgccatgt atgaactgtg ccaggggatg cgccagatca gccttcaatt cgtgcggctg
     3541 cagctgacct ttgaggagta ctccataatg aaggttttgc tgctactaag cacagttcca
     3601 aaagatggcc tcaagagcca ggctgcattt gaggagatga ggacaaatta catcaaagaa
     3661 ctgaggaaaa tggtcaccaa gtgtcccaac agttctggac agagttggca gaggttctac
     3721 caactgacca agcttctaga ctccatgcat gatctcgtga gtgacctgct agaattctgc
     3781 ttctacacct tccgagaatc ccaggctctg aaggtggagt ttcccgccat gctggtggag
     3841 atcatcaccg accagctgcc aaaggtggag tctgggaatg ccaaacccct ttactttcac
     3901 agaaagctcg aggactacaa agaccatgac ggtgattata aagatcatga catcgactac
     3961 aaggatgacg atgacaagta gtgagggccc gtttaaaccc gctgatcagc ctcgactgtg
     4021 ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa
     4081 ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt
     4141 aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa
     4201 gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc
     4261 agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt
     4321 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     4381 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     4441 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     4501 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     4561 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     4621 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     4681 aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag
     4741 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     4801 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     4861 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     4921 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     4981 aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     5041 gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcaag
     5101 agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg
     5161 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg
     5221 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc
     5281 tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga
     5341 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc
     5401 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag
     5461 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat
     5521 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg
     5581 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca
     5641 ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct
     5701 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg
     5761 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg
     5821 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc
     5881 gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat
     5941 gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta
     6001 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
     6061 ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta
     6121 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     6181 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag
     6241 ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     6301 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     6361 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     6421 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     6481 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     6541 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     6601 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     6661 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     6721 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     6781 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     6841 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     6901 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     6961 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     7021 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     7081 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     7141 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     7201 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     7261 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     7321 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     7381 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     7441 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     7501 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     7561 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     7621 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     7681 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     7741 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     7801 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     7861 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     7921 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     7981 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     8041 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     8101 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     8161 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     8221 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     8281 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     8341 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     8401 agtgccacct gacgtc
//
