LOCUS       V036715                 6946 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V036715
VERSION     V036715
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..2436
                     /label="CBLL1(NM_024814)"
                     /note="CBLL1(NM_024814)"
                     /gene="CBLL1"
     CDS             2443..2508
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    2546..2770
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2816..3244
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3258..3587
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3654..4448
                     /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    4622..4743
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4792..4808)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4816..4832
                     /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(4840..4870)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4885..4906
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5194..5779)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5950..6810)
                     /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(6811..6915)
                     /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 accatggatc acactgacaa tgagttacaa ggcactaata gttctggatc cttgggtggt
     1021 cttgatgttc gcagacgaat tcctataaag ctcatctcca aacaagcaaa caaagcgaaa
     1081 cctgcaccgc gaactcaaag aactataaac aggatgcctg caaaggctcc acctggtgat
     1141 gaagaaggat ttgattataa tgaagaagaa cggtatgact gtaaaggggg tgagctgttt
     1201 gcaaatcagc gaagatttcc tggacacctt ttttgggact ttcagataaa catcttaggt
     1261 gaaaaggatg atacaccagt tcatttctgt gacaagtgtg gattgcctat taaaatctat
     1321 gggagaatga ttccatgcaa gcatgttttt tgctatgact gtgctatttt acatgaaaaa
     1381 aagggagata agatgtgtcc aggctgtagt gatcctgtgc agcgaattga gcagtgtaca
     1441 cgaggttctc tcttcatgtg tagcattgtt caagggtgca agagaacata tttgtctcag
     1501 agagacttac aggctcatat caaccatcgc catatgagag ctggaaaacc tgttacccgt
     1561 gcttcacttg aaaatgttca tcctcctatt gccccaccac caactgaaat ccctgagcgt
     1621 tttataatgc caccagacaa gcaccatatg agccatattc cgccaaagca gcacatcatg
     1681 atgccaccac ctcctttgca acatgtgcca catgagcact ataatcagcc acatgaggat
     1741 attcgtgctc ctccagcaga attgtccatg gctccacctc cacctcgatc ggtcagtcag
     1801 gaaacctttc gtatttcaac aagaaaacac agcaatttaa taaccgtccc tattcaggat
     1861 gactcaaatt caggtgctag agaaccacca cctcctgccc cagcacctgc tcaccatcat
     1921 cctgaatatc agggtcaacc agtggtatcg caccctcatc atattatgcc tccacagcaa
     1981 cattatgcac cacccccacc tcctccacca ccaataagcc atccaatgcc acatcctccc
     2041 caggctgcag gtactcctca cttggtatat agccaagctc cacctccacc aatgacctct
     2101 gctccaccac caataacccc tccccctgga catattattg cccagatgcc accttatatg
     2161 aatcatcctc ctccaggacc tcccccacct caacatggtg gtccacctgt aactgcaccc
     2221 cctcctcacc attataatcc taactcatta ccccagttca ctgaagatca aggaactctg
     2281 agccctccat ttacacaacc agggggaatg agtcctggta tatggcctgc accaagaggg
     2341 ccaccaccac ctccacgatt gcagggtccg ccttctcaaa ccccacttcc tggaccacat
     2401 catccagatc agacaagata tagaccgtat taccaactcg aggactacaa agaccatgac
     2461 ggtgattata aagatcatga catcgactac aaggatgacg atgacaagta gtgagggccc
     2521 gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc
     2581 ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa
     2641 aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg
     2701 gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg
     2761 ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta tccccacgcg
     2821 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     2881 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     2941 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     3001 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     3061 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     3121 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     3181 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     3241 aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     3301 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     3361 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     3421 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     3481 atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat
     3541 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag
     3601 cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt cgcatgattg
     3661 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg
     3721 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg
     3781 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg
     3841 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg
     3901 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc
     3961 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc
     4021 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc
     4081 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc
     4141 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg
     4201 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct
     4261 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt
     4321 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc
     4381 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt
     4441 tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc
     4501 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg
     4561 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc
     4621 caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     4681 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     4741 ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct
     4801 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     4861 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     4921 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     4981 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     5041 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     5101 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     5161 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     5221 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     5281 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     5341 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     5401 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     5461 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     5521 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     5581 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     5641 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     5701 atccggcaaa caaaccaccg ctggtagcgg tttttttgtt tgcaagcagc agattacgcg
     5761 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     5821 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     5881 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     5941 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     6001 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     6061 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     6121 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     6181 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     6241 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     6301 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     6361 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     6421 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     6481 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     6541 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     6601 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     6661 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     6721 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     6781 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     6841 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     6901 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtc
//
