LOCUS       V027395                 8986 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V027395
VERSION     V027395
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"
     CDS             706..3645
                     /label="Nr3c2(NM_001083906)"
                     /note="Nr3c2(NM_001083906)"
                     /gene="Nr3c2"
     CDS             3652..3678
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             3682..3708
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             3712..3738
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     misc_feature    3762..4350
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4416..4464
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4666..5094
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5108..5437
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5504..6298
                     /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    6474..6595
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6644..6660)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6668..6684
                     /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(6692..6722)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6737..6758
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7046..7634)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7805..8665)
                     /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(8666..8770)
                     /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 gctctggcta gcgtttaaac ttaagcttgg
      661 taccgagctc ggatccacta gtccagtgtg gtggaattcg ccaccatgga aaccaaaggc
      721 taccacagtc tccctgaagg cctagatatg gaaaggcgct ggagtcaagt gtctcagacc
      781 ttggagcgtt cttctcttgg acctgcagag aggaccaatg agaacagcta catggagatt
      841 gtcaacgtca gctgcgtttc cggtgctact ccgaacaaca gtactcaagg gagcagcaaa
      901 gaaaaacacg aattactccc ttgtcttcag caagacaata gtcggtctgg gattttgcca
      961 tcagatatta aaactgagct ggaatccaag gaactttcag ccacggtggc tgagtccatg
     1021 ggtttataca tggattctgt gagagatgcc gagtacactt atgatcagca aaaccaacaa
     1081 ggaagcctga gcccggcaaa gatttatcaa aacatggagc agctggtgaa gttttacaaa
     1141 gagaatggtc acaggtcctc cacactgagt gctataagca ggcctttgag gtcattcatg
     1201 cctgactctg ggacctccat gaatggtggg gccttgcgtg ccatcgttaa gagcccaatc
     1261 atctgtcatg agaagagccc ctctgtttgc agcccgctca acatgccgtc ttcagtatgc
     1321 agccccgcgg gcatcaactc catgtcctcc tccacagcta gctttggcag tttcccagtg
     1381 cacagtccca tcactcaagg aacctcactg acatgctccc ccagtgttga aaatagaggc
     1441 tcaaggtcac acagccccgt acatgcgagc aatgtgggct ctcctctttc aagtccatta
     1501 agcagcatga aatccccaat ttccagccct ccaagtcact gcagtgtaaa atctccagta
     1561 tccagtccaa acaatgtccc tctgcgctcc tctgtgtcaa gcccggcaaa tcttaacaat
     1621 tcaaggtgct ctgtttccag cccttccaac accaacaata gatctacact ctccagcccg
     1681 acagctagca cagtggggtc cattggcagc cccatcagca atgccttcag ctataccact
     1741 tcaggcgctt cggctggagc cggtgccatc caggatatgg ttcccagtcc agacacccac
     1801 gagaaaggtg ctcacgacgt tcctttccct aagacagagg aagtcgagaa ggccatttcc
     1861 aatggtgtga ctggtcagct caacattgtc cagtacataa aaccagaacc agatggggct
     1921 ttcagcagtt cctgcctagg aggaaacaac aaaatcaacc ccagttctcc gttctctgta
     1981 ccaataaagc aagagtcaag caagcactca tgttcaggcg cctcttttaa agggaacccc
     2041 acagtcaacc catttccatt catggatggc tcgtactttt cttttatgga tgataaagac
     2101 tattattccc tatcaggaat cttaggacca cctgtgcccg gctttgatag tagctgtgaa
     2161 ggcagtgcgt tcccgggggg gattaagcaa gaaccagatg atgggagcta tttccctgaa
     2221 accagcatcc catcatctgc catcattggt gtgaattcag gtggacagtc ctttcactac
     2281 cggattggtg ctcaaggtac aatatcttta tcacggtcac ctagagacca atctttccaa
     2341 cacttgagtt cctttccgcc tgtcaatgca ttagtggagt catggaaacc acacggtgac
     2401 ctgtcatcta ggagaagtga tgggtacccg gtcctagagt acattccaga aaacgtgtca
     2461 agctctactt tacgaagtgt ttctactgga tcctcaagac cttccaagat ctgcttggtg
     2521 tgtggagatg aggcttctgg gtgtcactat ggggtagtga cctgcggcag ctgcaaagtc
     2581 ttcttcaaaa gagccgtgga aggacaacac aactatctgt gtgctggaag aaatgactgc
     2641 attattgata agattcggag aaagaactgt cctgcctgca ggctccagaa atgcctccaa
     2701 gccggcatga acttaggagc tcgaaagtca aagaagctgg ggaagttaaa aggccttcac
     2761 gaggagcagc cacagcagcc cccaccgcca ccaccccaga gcccagaaga ggggaccaca
     2821 tacattgctc ctaccaagga gccatcagtg aactctgcgc tggtcccgca gctcgcctcg
     2881 atcacgcgtg cgctcacgcc atccccgtcc atgatcctgg agaacatcga gcccgagatc
     2941 gtgtatgcag gctacgacaa ttccaagcct gacaccgccg agagcctgct ctccacgctc
     3001 aaccgcctgg cgggcaagca gatgatccaa gtcgtgaagt gggccaaggt acttccagga
     3061 tttaaaaact tgcctcttga ggaccaaatt accctcatcc agtattcttg gatgtgtcta
     3121 tcatcgtttg ccttgagttg gagatcgtac aaacatacga acagccaatt tctctatttt
     3181 gcaccagacc tagtttttaa tgaagagaag atgcatcagt ctgccatgta tgagctgtgc
     3241 caggggatgc ggcagatcag ccttcagttc gtgcggcttc agctgacctt tgaggagtac
     3301 tccataatga aggttttgct gctactgagc acagttccaa aagatggcct caagagccag
     3361 gctgcgtttg aggagatgag gacaaattac atcaaagaac tgaggaaaat ggtcaccaag
     3421 tgtcccaaca gttctggaca gagttggcag aggttctacc aactgacgaa gcttctagac
     3481 tccatgcatg atttggtgaa tgacctgcta gaattctgct tctatacctt ccgagaatcc
     3541 caggctctga aggtggagtt tcccgccatg ctggtggaga tcatcagcga ccagctgcca
     3601 aaggtggaat ccgggaacgc caagcccctt tactttcaca ggaagctcga gtacccatac
     3661 gacgtacctg attatgcaac ttatccttat gatgtacctg attatgctag ctatccttat
     3721 gatgtcccag actacgcata gtgagggccc gatatctcga caatcaacct ctggattaca
     3781 aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat
     3841 acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct
     3901 ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac
     3961 gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca
     4021 cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca
     4081 tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg
     4141 tggtgttgtc ggggaagctg acgtcctttc catggctgct cgcctgtgtt gccacctgga
     4201 ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt
     4261 cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga
     4321 gtcggatctc cctttgggcc gcctccccgc ctggaaacgg gggaggctaa ctgaaacacg
     4381 gaaggagaca ataccggaag gaacccgcgc tatgacggca ataaaaagac agaataaaac
     4441 gcacgggtgt tgggtcgttt gttcataaac gcggggttcg gtcccagggc tggcactctg
     4501 tcgatacccc accgagaccc cattggggcc aatacgcccg cgtttcttcc ttttccccac
     4561 cccacccccc aagttcgggt gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc
     4621 ctgccatagc agatctgcgc agctggggct ctagggggta tccccacgcg ccctgtagcg
     4681 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     4741 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     4801 cccgtcaagc tctaaatcgg ggcatccctt tagggttccg atttagtgct ttacggcacc
     4861 tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga
     4921 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     4981 ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg attttgggga
     5041 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct
     5101 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     5161 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc
     5221 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac
     5281 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     5341 aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta
     5401 gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc
     5461 cattttcgga tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg
     5521 attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca
     5581 acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt
     5641 tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg
     5701 gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga
     5761 agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca
     5821 ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct
     5881 tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac
     5941 tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc
     6001 gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt
     6061 gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt
     6121 catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg
     6181 tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat
     6241 cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc
     6301 gggactctgg ggttcgcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt
     6361 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
     6421 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt
     6481 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     6541 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     6601 tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg
     6661 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     6721 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     6781 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     6841 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     6901 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     6961 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     7021 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     7081 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     7141 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     7201 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca atgctcacgc tgtaggtatc
     7261 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     7321 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     7381 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     7441 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta
     7501 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     7561 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     7621 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     7681 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     7741 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     7801 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     7861 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     7921 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     7981 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     8041 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     8101 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     8161 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     8221 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     8281 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     8341 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     8401 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     8461 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     8521 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     8581 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     8641 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     8701 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     8761 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc
     8821 cgatccccta tggtcgactc tcagtacaat ctgctctgat gccgcatagt taagccagta
     8881 tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac
     8941 aacaaggcaa ggcttgaccg acaattgcat gaagaatctg cttagg
//
