LOCUS       V028552                 6923 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V028552
VERSION     V028552
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             941..2413
                     /label="Stk33(NM_054103)"
                     /note="Stk33(NM_054103)"
                     /gene="Stk33"
     CDS             2420..2485
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    2523..2747
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2793..3221
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3235..3564
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3631..4425
                     /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    4599..4720
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4769..4785)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4793..4809
                     /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(4817..4847)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4862..4883
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5171..5756)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5927..6787)
                     /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(6788..6892)
                     /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 atccgccacc atggctgacc ccagcttgaa
      961 tgacaaccct acagcatgcc ctcactgtgc atcctctcag gctggcctac tgtgtgtatg
     1021 tccagcaggc aagtctccag tcctggtggt ggaaatgtca cagacatcga gtattggtag
     1081 tacagaattt tttgcttcac aagaaagaaa aaaggaaaga aataccagca gagaatcttc
     1141 tctaaaagat ttgtccataa gaacttcaaa tgtggagaga aaacctcagg cacaatggag
     1201 tcggagcaat gtcacagtag gaaaaatccc acacataaga atggacgatg gagcaggtat
     1261 cgaggaattc tatacctttg gaagaatatt gggacagggg agctttggaa tggtctttga
     1321 agctatagac aaggaaacag gagctaagtg ggcaattaaa aaagtgaata aagaaaaggc
     1381 tggaagttct gcaatgaagc tactggagcg ggaggtgagc atcctgaaga ctgtcaacca
     1441 tcaacacatc atccacctgg aacaagtgtt tgagtcgcct cagaaaatgt atctcgtgat
     1501 ggagctttgt gaggatggag aactcaaagc agttatggat caaagagggc acttctcaga
     1561 gaacgagaca aggctgataa ttcaaagtct tgcatcagcc atcgcatatc ttcataacaa
     1621 ggatatagtg cacagagatc taaagctgga aaacataatg gttaaaagca gctttataga
     1681 tgataacaat gaaatgaact taaacataaa ggtgactgat tttggcttgt ctgtgcagaa
     1741 gcatggctcc aggagtgaag gcatgatgca gactacatgt gggactccta tctatatggc
     1801 accagaggtc atcaatgccc atgactacag ccagcagtgt gacatttgga gcataggtgt
     1861 cataatgttc attttactgt gtggagagcc accctttttg gcaaattcag aagaaaagct
     1921 ctatgaatta ataaaaaagg gagaactacg atttgaaaat ccagtctggg aatctgtaag
     1981 tgattctgca aaaaatactt tgaaacaact catgaaagta gatcctgctc acagaatcac
     2041 agctaaggaa cttctagata accaatggtt gacaggcaat accctttctt cagcaagacc
     2101 aaccaatgta ttagaaatga tgaaagaatg gaaaaataac ccagaaagtg atgaggagac
     2161 caacacagat gaggagactg agcagagcgc tgtctacagt ccatctgcaa acacagcaaa
     2221 gcagcccacc aatgcagcca agaagcctgc tgcagagagt gttggcatga cctcttcaaa
     2281 ctcatcgtcc agcaaactcc tgtctgctga aagcaaagca gaaccagaga aaagctccga
     2341 gactgtaggc catgcatcag tggctaaaac cactctgaaa tccactacct tgtttcgagg
     2401 caagaaaagg ctcctcgagg actacaaaga ccatgacggt gattataaag atcatgacat
     2461 cgactacaag gatgacgatg acaagtagtg agggcccgtt taaacccgct gatcagcctc
     2521 gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac
     2581 cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg
     2641 tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga
     2701 ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga
     2761 aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc
     2821 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     2881 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     2941 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     3001 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     3061 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     3121 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg
     3181 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt
     3241 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     3301 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     3361 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     3421 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     3481 tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt
     3541 tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct
     3601 gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt
     3661 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc
     3721 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag
     3781 accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg
     3841 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac
     3901 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc
     3961 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc
     4021 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc
     4081 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg
     4141 ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat
     4201 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc
     4261 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa
     4321 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat
     4381 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt
     4441 tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg
     4501 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc
     4561 agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata
     4621 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     4681 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga
     4741 cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     4801 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     4861 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     4921 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     4981 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     5041 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     5101 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     5161 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     5221 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     5281 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     5341 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     5401 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     5461 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     5521 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     5581 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     5641 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     5701 gtagcggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     5761 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     5821 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     5881 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     5941 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     6001 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     6061 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     6121 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     6181 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     6241 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     6301 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     6361 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     6421 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     6481 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     6541 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     6601 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     6661 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     6721 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     6781 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     6841 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     6901 cccgaaaagt gccacctgac gtc
//
