LOCUS       V022830                 7238 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V022830
VERSION     V022830
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             941..2728
                     /label="Nr2c2(NM_011630)"
                     /note="Nr2c2(NM_011630)"
                     /gene="Nr2c2"
     CDS             2735..2800
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    2838..3062
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3108..3536
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3550..3879
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3946..4740
                     /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    4914..5035
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5084..5100)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5108..5124
                     /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(5132..5162)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5177..5198
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5486..6071)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6242..7102)
                     /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(7103..7207)
                     /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 atccgccacc atgaccagcc cctccccgcg
      961 catccagata atttccaccg actctgcggt agcctcacct cagcgcattc agattgtaac
     1021 agaccagcag acaggacaaa agatccagat agtcaccgca gtggatgcct ctggatcctc
     1081 taaacagcag ttcatcctaa ccagcccaga tggagctgga actgggaagg tgatcctggc
     1141 ttctccggaa acatccagtg ccaagcagct catattcacc acctcggaca accttgtccc
     1201 tggcaggatc cagatcgtca cggattctgc ttctgtggag cgtttgctgg ggaaggcaga
     1261 cgtccagcgg ccccaggtgg tggagtactg tgtggtctgt ggcgacaaag cctctggccg
     1321 gcactatggg gctgtcagtt gtgaaggttg caaaggtttc ttcaaaagga gcgtgaggaa
     1381 gaatctgacc tacagctgtc ggagcagcca agactgcatc atcaacaagc accaccgtaa
     1441 ccgctgccag ttctgccggc tgaagaagtg cctggagatg ggcatgaaaa tggagtctgt
     1501 acagagtgaa cggaagccct ttgatgtgca acgggagaaa ccaagcaatt gtgctgcttc
     1561 cactgagaag atctatatcc ggaaagacct gagaagtcct ctgatagcca ctcccacatt
     1621 tgtggcagac aaagatggag caagacaaac aggtcttctt gatccaggga tgcttgtgaa
     1681 catccaacag cctttgatac gtgaggatgg tacagttctc ctggccgcgg attccaaggc
     1741 tgaaacaagc caaggagctc taggtacact ggcaaatgta gtgacctctt tggccaacct
     1801 gagtgaatct ttgaacaacg gtgatgcttc agaaatgcag ccagaggacc agtctgcaag
     1861 tgagattact cgggcatttg acaccttagc gaaagcactt aataccacag atagtgcttc
     1921 acctccaagc ctggcagatg ggatagatgc tagtggagga gggagtatcc atgtcatcag
     1981 cagagatcag tcaacaccca tcattgaagt tgaaggccct ctcctttcag acacacatgt
     2041 cacattcaag cttacaatgc ccagtcctat gccagagtac ctcaatgtac attacatctg
     2101 tgagtctgca tcccgcctgc ttttcctctc catgcactgg gcaaggtcaa tcccagcctt
     2161 ccaggcactt ggacaggact gtaataccag cctggtgagg gcctgctgga atgagctctt
     2221 cactcttggc ctggcccagt gtgcccaggt catgagtctc tccaccatcc tggcagccat
     2281 tgtcaaccac ctacagaaca gcatccagga agataagctt tctggtgacc ggataaagca
     2341 agtgatggag cacatctgga agctgcagga gttctgtaac agcatggcga aactggatat
     2401 agacggctat gagtacgcat accttaaagc tatagttctc tttagtcccg atcatccagg
     2461 tttgacaggc acaagccaga ttgagaaatt tcaggagaag gcacagatgg aattacagga
     2521 ctatgtgcag aaaacctact ctgaagatac ttacagattg gccaggattc ttgtccgcct
     2581 accagcactc aggctcatga gctccaacat aacagaagaa ctttttttta ctggtctcat
     2641 tggcaatgtt tcaatagaca gcataattcc ttacatcctc aagatggaga cagcagaata
     2701 taatggccag atcactggag ccagtctact cgaggactac aaagaccatg acggtgatta
     2761 taaagatcat gacatcgact acaaggatga cgatgacaag tagtgagggc ccgtttaaac
     2821 ccgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc
     2881 cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga
     2941 aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga
     3001 cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat
     3061 ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag
     3121 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     3181 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     3241 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     3301 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     3361 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     3421 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
     3481 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt
     3541 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     3601 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     3661 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     3721 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     3781 ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag
     3841 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata
     3901 tccattttcg gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat
     3961 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     4021 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     4081 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg
     4141 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     4201 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     4261 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     4321 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     4381 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     4441 gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc
     4501 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     4561 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     4621 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     4681 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     4741 gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt
     4801 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
     4861 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt
     4921 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     4981 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     5041 tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg
     5101 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     5161 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     5221 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     5281 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     5341 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     5401 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     5461 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     5521 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     5581 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     5641 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     5701 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     5761 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     5821 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     5881 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     5941 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     6001 aacaaaccac cgctggtagc ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     6061 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     6121 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     6181 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     6241 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     6301 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     6361 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     6421 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     6481 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     6541 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     6601 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     6661 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     6721 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     6781 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     6841 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     6901 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     6961 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     7021 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     7081 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     7141 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     7201 ttccgcgcac atttccccga aaagtgccac ctgacgtc
//
