LOCUS       V032534                 7116 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V032534
VERSION     V032534
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"
     promoter        678..696
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             756..1796
                     /label="JUNB(NM_002229)"
                     /note="JUNB(NM_002229)"
                     /gene="JUNB"
     CDS             1803..1868
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    1892..2480
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2546..2594
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      2796..3224
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3238..3567
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3634..4428
                     /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    4604..4725
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4774..4790)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4798..4814
                     /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(4822..4852)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4867..4888
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5176..5764)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5935..6795)
                     /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(6796..6900)
                     /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 gctctctggc taactagaga acccactgct
      661 tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagcgttta
      721 aacttaagct tggtaccgag ctcgaattcg ccaccatgtg cactaaaatg gaacagccct
      781 tctaccacga cgactcatac acagctacgg gatacggccg ggcccctggt ggcctctctc
      841 tacacgacta caaactcctg aaaccgagcc tggcggtcaa cctggccgac ccctaccgga
      901 gtctcaaagc gcctggggct cgcggacccg gcccagaggg cggcggtggc ggcagctact
      961 tttctggtca gggctcggac accggcgcgt ctctcaagct cgcctcttcg gagctggaac
     1021 gcctgattgt ccccaacagc aacggcgtga tcacgacgac gcctacaccc ccgggacagt
     1081 acttttaccc ccgcgggggt ggcagcggtg gaggtgcagg gggcgcaggg ggcggcgtca
     1141 ccgaggagca ggagggcttc gccgacggct ttgtcaaagc cctggacgat ctgcacaaga
     1201 tgaaccacgt gacacccccc aacgtgtccc tgggcgctac cggggggccc ccggctgggc
     1261 ccgggggcgt ctacgccggc ccggagccac ctcccgttta caccaacctc agcagctact
     1321 ccccagcctc tgcgtcctcg ggaggcgccg gggctgccgt cgggaccggg agctcgtacc
     1381 cgacgaccac catcagctac ctcccacacg cgccgccctt cgccggtggc cacccggcgc
     1441 agctgggctt gggccgcggc gcctccacct tcaaggagga accgcagacc gtgccggagg
     1501 cgcgcagccg ggacgccacg ccgccggtgt cccccatcaa catggaagac caagagcgca
     1561 tcaaagtgga gcgcaagcgg ctgcggaacc ggctggcggc caccaagtgc cggaagcgga
     1621 agctggagcg catcgcgcgc ctggaggaca aggtgaagac gctcaaggcc gagaacgcgg
     1681 ggctgtcgag taccgccggc ctcctccggg agcaggtggc ccagctcaaa cagaaggtca
     1741 tgacccacgt cagcaacggc tgtcagctgc tgcttggggt caagggacac gccttcggat
     1801 ccgactacaa agaccatgac ggtgattata aagatcatga catcgactac aaggatgacg
     1861 atgacaagta gtgagggccc gatatctcga caatcaacct ctggattaca aaatttgtga
     1921 aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt
     1981 aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa
     2041 atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt
     2101 gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct
     2161 cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg
     2221 ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc
     2281 ggggaagctg acgtcctttc catggctgct cgcctgtgtt gccacctgga ttctgcgcgg
     2341 gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct
     2401 gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc
     2461 cctttgggcc gcctccccgc ctggaaacgg gggaggctaa ctgaaacacg gaaggagaca
     2521 ataccggaag gaacccgcgc tatgacggca ataaaaagac agaataaaac gcacgggtgt
     2581 tgggtcgttt gttcataaac gcggggttcg gtcccagggc tggcactctg tcgatacccc
     2641 accgagaccc cattggggcc aatacgcccg cgtttcttcc ttttccccac cccacccccc
     2701 aagttcgggt gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc
     2761 agatctgcgc agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag
     2821 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     2881 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     2941 tctaaatcgg ggcatccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     3001 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     3061 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     3121 actcaaccct atctcggtct attcttttga tttataaggg attttgggga tttcggccta
     3181 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg
     3241 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     3301 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
     3361 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
     3421 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     3481 atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc
     3541 ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga
     3601 tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca
     3661 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc
     3721 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc
     3781 aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg
     3841 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg
     3901 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct
     3961 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct
     4021 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa
     4081 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa
     4141 ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc
     4201 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt
     4261 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct
     4321 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc
     4381 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg
     4441 ggttcgcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac
     4501 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat
     4561 cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc
     4621 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     4681 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc
     4741 gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     4801 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     4861 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     4921 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     4981 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     5041 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     5101 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     5161 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     5221 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     5281 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     5341 tctcccttcg ggaagcgtgg cgctttctca atgctcacgc tgtaggtatc tcagttcggt
     5401 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     5461 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     5521 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     5581 cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct
     5641 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     5701 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     5761 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     5821 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     5881 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     5941 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     6001 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     6061 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     6121 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     6181 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     6241 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     6301 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     6361 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     6421 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     6481 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     6541 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     6601 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     6661 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     6721 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     6781 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     6841 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     6901 cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc cgatccccta
     6961 tggtcgactc tcagtacaat ctgctctgat gccgcatagt taagccagta tctgctccct
     7021 gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa
     7081 ggcttgaccg acaattgcat gaagaatctg cttagg
//
