LOCUS       V046021                 7708 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V046021
VERSION     V046021
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"
     protein_bind    820..838
                     /bound_moiety="tetracycline repressor TetR"
                     /gene="tetO"
                     /note="bacterial operator O2 for the tetR and tetA genes"
                     /label="tet operator"
     protein_bind    841..859
                     /bound_moiety="tetracycline repressor TetR"
                     /gene="tetO"
                     /note="bacterial operator O2 for the tetR and tetA genes"
                     /label="tet operator"
     CDS             990..3530
                     /label="MAP3K11(NM_002419)"
                     /note="MAP3K11(NM_002419)"
                     /gene="MAP3K11"
     polyA_signal    3666..3890
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     protein_bind    4174..4221
                     /bound_moiety="FLP recombinase from the Saccharomyces
                     cerevisiae 2μ plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core
                     sequence TCTAGAAA (Turan and Bode, 2011)."
                     /label="FRT"
     CDS             4229..5251
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /note="confers resistance to hygromycin"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /transl_table=1
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     *"
                     /label="HygR"
     polyA_signal    5381..5502
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5551..5567)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5575..5591
                     /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(5599..5629)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5644..5665
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5953..6541)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6712..7572)
                     /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(7573..7677)
                     /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 ccctatcagt gatagagatc
      841 tccctatcag tgatagagat cgtcgacgag ctcgtttagt gaaccgtcag atcgcctgga
      901 gacgccatcc acgctgtttt gacctccata gaagacaccg ggaccgatcc agcctccgga
      961 ctctagcgtt taaacttaag cttgccacca tggagccctt gaagagcctc ttcctcaaga
     1021 gccctctagg gtcatggaat ggcagtggca gcgggggtgg tgggggcggt ggaggaggcc
     1081 ggcctgaggg gtctccaaag gcagcgggtt atgccaaccc ggtgtggaca gccctgttcg
     1141 actacgagcc cagtgggcag gatgagctgg ccctgaggaa gggtgaccgt gtggaggtgc
     1201 tgtcccggga cgcagccatc tcaggagacg agggctggtg ggcgggccag gtgggtggcc
     1261 aggtgggcat cttcccgtcc aactatgtgt ctcggggtgg cggcccgccc ccctgcgagg
     1321 tggccagctt ccaggagctg cggctggagg aggtgatcgg cattggaggc tttggcaagg
     1381 tgtacagggg cagctggcga ggtgagctgg tggctgtgaa ggcagctcgc caggaccccg
     1441 atgaggacat cagtgtgaca gccgagagcg ttcgccagga ggcccggctc ttcgccatgc
     1501 tggcacaccc caacatcatt gccctcaagg ctgtgtgcct ggaggagccc aacctgtgcc
     1561 tggtgatgga gtatgcagcc ggtgggcccc tcagccgagc tctggccggg cggcgcgtgc
     1621 ctccccatgt gctggtcaac tgggctgtgc agattgcccg tgggatgcac tacctgcact
     1681 gcgaggccct ggtgcccgtc atccaccgtg atctcaagtc caacaacatt ttgctgctgc
     1741 agcccattga gagtgacgac atggagcaca agaccctgaa gatcaccgac tttggcctgg
     1801 cccgagagtg gcacaaaacc acacaaatga gtgccgcggg cacctacgcc tggatggctc
     1861 ctgaggttat caaggcctcc accttctcta agggcagtga cgtctggagt tttggggtgc
     1921 tgctgtggga actgctgacc ggggaggtgc cataccgtgg cattgactgc cttgctgtgg
     1981 cctatggcgt agctgttaac aagctcacac tgcccatccc atccacctgc cccgagccct
     2041 tcgcacagct tatggccgac tgctgggcgc aggaccccca ccgcaggccc gacttcgcct
     2101 ccatcctgca gcagttggag gcgctggagg cacaggtcct acgggaaatg ccgcgggact
     2161 ccttccattc catgcaggaa ggctggaagc gcgagatcca gggtctcttc gacgagctgc
     2221 gagccaagga aaaggaacta ctgagccgcg aggaggagct gacgcgagcg gcgcgcgagc
     2281 agcggtcaca ggcggagcag ctgcggcggc gcgagcacct gctggcccag tgggagctag
     2341 aggtgttcga gcgcgagctg acgctgctgc tgcagcaggt ggaccgcgag cgaccgcacg
     2401 tgcgccgccg ccgcgggaca ttcaagcgca gcaagctccg ggcgcgcgac ggcggcgagc
     2461 gtatcagcat gccactcgac ttcaagcacc gcatcaccgt gcaggcctca cccggccttg
     2521 accggaggag aaacgtcttc gaggtcgggc ctggggattc gcccaccttt ccccggttcc
     2581 gagccatcca gttggagcct gcagagccag gccaggcatg gggccgccag tccccccgac
     2641 gtctggagga ctcaagcaat ggagagcggc gagcatgctg ggcttggggt cccagttccc
     2701 ccaagcctgg ggaagcccag aatgggagga gaaggtcccg catggacgaa gccacatggt
     2761 acctggattc agatgactca tcccccttag gatctccttc cacaccccca gcactcaatg
     2821 gtaacccccc gcggcctagc ctggagcccg aggagcccaa gaggcctgtc cccgcagagc
     2881 gcggtagcag ctctgggacg cccaagctga tccagcgggc gctgctgcgc ggcaccgccc
     2941 tgctcgcctc gctgggcctt ggccgcgacc tgcagccgcc gggaggccca ggacgcgagc
     3001 gcggggagtc cccgacaaca ccccccacgc caacgcccgc gccctgcccg accgagccgc
     3061 ccccttcccc gctcatctgc ttctcgctca agacgcccga ctccccgccc actcctgcac
     3121 ccctgttgct ggacctgggt atccctgtgg gccagcggtc agccaagagc ccccgacgtg
     3181 aggaggagcc ccgcggaggc actgtctcac ccccaccggg gacatcacgc tctgctcctg
     3241 gcaccccagg caccccacgt tcaccacccc tgggcctcat cagccgacct cggccctcgc
     3301 cccttcgcag ccgcattgat ccctggagct ttgtgtcagc tgggccacgg ccttctcccc
     3361 tgccatcacc acagcctgca ccccgccgag caccctggac cttgttcccg gactcagacc
     3421 ccttctggga ctccccacct gccaacccct tccagggggg cccccaggac tgcagggcac
     3481 agaccaaaga catgggtgcc caggccccgt gggtgccgga agcggggcct gtgagcggct
     3541 ggcggctgtt caagaagatt agctgaggat ccactagtcc agtgtggtgg aattctgcag
     3601 atatccagca cagtggcggc cgctcgagtc tagagggccc gtttaaaccc gctgatcagc
     3661 ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt
     3721 gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca
     3781 ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga
     3841 ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc
     3901 ggaaagaacc agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag
     3961 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     4021 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     4081 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     4141 aaaacttgat tagggtgatg gttcacgtac ctagaagttc ctattccgaa gttcctattc
     4201 tctagaaagt ataggaactt ccttggccaa aaagcctgaa ctcaccgcga cgtctgtcga
     4261 gaagtttctg atcgaaaagt tcgacagcgt ctccgacctg atgcagctct cggagggcga
     4321 agaatctcgt gctttcagct tcgatgtagg agggcgtgga tatgtcctgc gggtaaatag
     4381 ctgcgccgat ggtttctaca aagatcgtta tgtttatcgg cactttgcat cggccgcgct
     4441 cccgattccg gaagtgcttg acattgggga attcagcgag agcctgacct attgcatctc
     4501 ccgccgtgca cagggtgtca cgttgcaaga cctgcctgaa accgaactgc ccgctgttct
     4561 gcagccggtc gcggaggcca tggatgcgat cgctgcggcc gatcttagcc agacgagcgg
     4621 gttcggccca ttcggaccgc aaggaatcgg tcaatacact acatggcgtg atttcatatg
     4681 cgcgattgct gatccccatg tgtatcactg gcaaactgtg atggacgaca ccgtcagtgc
     4741 gtccgtcgcg caggctctcg atgagctgat gctttgggcc gaggactgcc ccgaagtccg
     4801 gcacctcgtg cacgcggatt tcggctccaa caatgtcctg acggacaatg gccgcataac
     4861 agcggtcatt gactggagcg aggcgatgtt cggggattcc caatacgagg tcgccaacat
     4921 cttcttctgg aggccgtggt tggcttgtat ggagcagcag acgcgctact tcgagcggag
     4981 gcatccggag cttgcaggat cgccgcggct ccgggcgtat atgctccgca ttggtcttga
     5041 ccaactctat cagagcttgg ttgacggcaa tttcgatgat gcagcttggg cgcagggtcg
     5101 atgcgacgca atcgtccgat ccggagccgg gactgtcggg cgtacacaaa tcgcccgcag
     5161 aagcgcggcc gtctggaccg atggctgtgt agaagtactc gccgatagtg gaaaccgacg
     5221 ccccagcact cgtccgaggg caaaggaata gcacgtacta cgagatttcg attccaccgc
     5281 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct
     5341 ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta
     5401 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     5461 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc
     5521 gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     5581 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     5641 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     5701 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     5761 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     5821 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     5881 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     5941 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     6001 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     6061 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     6121 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     6181 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     6241 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     6301 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     6361 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     6421 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     6481 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     6541 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     6601 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     6661 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     6721 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     6781 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     6841 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     6901 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     6961 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     7021 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     7081 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     7141 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     7201 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     7261 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     7321 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     7381 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     7441 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     7501 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     7561 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     7621 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     7681 atttccccga aaagtgccac ctgacgtc
//
