LOCUS       V029741                 7326 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V029741
VERSION     V029741
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..2006
                     /label="CBLIF(NM_005142)"
                     /note="CBLIF(NM_005142)"
                     /gene="CBLIF"
     CDS             2013..2078
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2102..2690
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2756..2804
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3006..3434
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3448..3777
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3844..4638
                     /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    4814..4935
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4984..5000)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5008..5024
                     /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(5032..5062)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5077..5098
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5386..5974)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6145..7005)
                     /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(7006..7110)
                     /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 ccaccatggc ctggtttgcc ctctacctcc
      781 tgagccttct ctgggctaca gctgggacta gtacccagac ccagagttca tgctccgttc
      841 cctcagcaca ggagcccttg gtcaatggaa tacaagtact catggagaac tcggtgactt
      901 catcagccta cccaaacccc agcatcctga ttgccatgaa tctggccgga gcctacaact
      961 tgaaggccca gaagctcctg acttaccagc tcatgtccag cgacaacaac gatctaacca
     1021 ttgggcagct cggcctcacc atcatggccc tcacctcctc ctgccgagac cctggggata
     1081 aagtatccat tctacaaaga caaatggaga actgggcacc ttccagcccc aacgctgaag
     1141 catcagcctt ctatgggccc agtctagcga tcttggcact gtgccagaag aactctgagg
     1201 cgaccttgcc gatagccgtc cgctttgcca agaccctgct ggccaactcc tctcccttca
     1261 atgtagacac aggagcaatg gcaaccttgg ctctgacctg tatgtacaac aagatccctg
     1321 taggttcaga ggaaggttac agatccctgt ttggtcaggt actaaaggat attgtggaga
     1381 aaatcagcat gaagatcaaa gataatggca tcattggaga catctacagt actggcctcg
     1441 ccatgcaggc tctctctgta acacctgagc catctaaaaa ggaatggaac tgcaagaaga
     1501 ctacggatat gatactcaat gagattaagc aggggaaatt ccacaacccc atgtccattg
     1561 ctcaaatcct cccttccctg aaaggcaaga catacctaga tgtgccccag gtcacttgta
     1621 gtcctgatca tgaggtacaa ccaactctac ccagcaaccc tggccctggc cccacctctg
     1681 catctaacat cactgtcata tacaccataa ataaccagct gaggggggtt gagctgctct
     1741 tcaacgagac catcaatgtt agtgtgaaaa gtgggtcagt gttacttgtt gtcctagagg
     1801 aagcacagcg caaaaatcct atgttcaaat ttgaaaccac aatgacatct tggggccttg
     1861 tcgtctcttc tatcaacaat atcgcggaaa atgttaatca caagacatac tggcagtttc
     1921 ttagtggtgt aacacctttg aatgaagggg ttgctgacta catacccttc aaccacgagc
     1981 acatcacagc caatttcaca cagtacggat ccgactacaa agaccatgac ggtgattata
     2041 aagatcatga catcgactac aaggatgacg atgacaagta gtgagggccc gatatctcga
     2101 caatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc
     2161 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg
     2221 tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt
     2281 gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac
     2341 tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc
     2401 tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct
     2461 gttgggcact gacaattccg tggtgttgtc ggggaagctg acgtcctttc catggctgct
     2521 cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct
     2581 caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct
     2641 tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc ctggaaacgg
     2701 gggaggctaa ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca
     2761 ataaaaagac agaataaaac gcacgggtgt tgggtcgttt gttcataaac gcggggttcg
     2821 gtcccagggc tggcactctg tcgatacccc accgagaccc cattggggcc aatacgcccg
     2881 cgtttcttcc ttttccccac cccacccccc aagttcgggt gaaggcccag ggctcgcagc
     2941 caacgtcggg gcggcaggcc ctgccatagc agatctgcgc agctggggct ctagggggta
     3001 tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     3061 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     3121 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg ggcatccctt tagggttccg
     3181 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
     3241 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     3301 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
     3361 tttataaggg attttgggga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     3421 atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc
     3481 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga
     3541 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     3601 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     3661 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc
     3721 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag
     3781 ctcccgggag cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt
     3841 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     3901 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
     3961 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     4021 ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     4081 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     4141 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     4201 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     4261 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     4321 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc
     4381 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     4441 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     4501 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     4561 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     4621 cttgacgagt tcttctgagc gggactctgg ggttcgcgaa atgaccgacc aagcgacgcc
     4681 caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg
     4741 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt
     4801 cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     4861 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     4921 catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc
     4981 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     5041 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     5101 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     5161 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     5221 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     5281 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     5341 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     5401 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     5461 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     5521 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     5581 atgctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     5641 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     5701 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     5761 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     5821 tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     5881 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     5941 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     6001 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     6061 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     6121 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     6181 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     6241 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     6301 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     6361 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     6421 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     6481 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     6541 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     6601 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     6661 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     6721 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     6781 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     6841 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     6901 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     6961 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     7021 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     7081 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     7141 cgacggatcg ggagatctcc cgatccccta tggtcgactc tcagtacaat ctgctctgat
     7201 gccgcatagt taagccagta tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc
     7261 gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg acaattgcat gaagaatctg
     7321 cttagg
//
