LOCUS       V035667                 8818 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V035667
VERSION     V035667
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        202..581
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        582..785
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        830..848
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             874..1590
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /transl_table=1
                     /translation="M,V,SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKL
                     TLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFK
                     DDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNG
                     IKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMV
                     LLEFVTAAGITLGMDELYK"
                     /label="EGFP"
     CDS             1642..4308
                     /label="PKN3(NM_013355)"
                     /note="PKN3(NM_013355)"
                     /gene="PKN3"
     polyA_signal    4385..4609
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4655..5083
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5097..5426
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5493..6287
                     /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    6461..6582
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6631..6647)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6655..6671
                     /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(6679..6709)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6724..6745
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7033..7618)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7789..8649)
                     /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(8650..8754)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
       61 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
      121 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg
      181 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta
      241 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      301 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      361 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      421 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
      481 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
      541 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      601 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      661 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
      721 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      781 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc
      841 actataggga gacccaagct ggctagcgcc accatggtga gcaagggcga ggagctgttc
      901 accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc
      961 gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc
     1021 accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg
     1081 cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg
     1141 cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc
     1201 cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc
     1261 gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac
     1321 aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc
     1381 cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc
     1441 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc cgccctgagc
     1501 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg
     1561 atcactctcg gcatggacga gctgtacaag ggaggcggag gtagtggagg agggggaagc
     1621 ggaggcggag ggagtgaatt catggaggag ggggcgccgc ggcagcctgg gccgagccag
     1681 tggcccccag aggatgagaa ggaggtgatc cgccgggcca tccagaaaga gctgaagatc
     1741 aaggaggggg tggagaacct gcggcgcgtg gccacagacc gccgccactt gggccatgtg
     1801 cagcagctgc tgcggtcctc caaccgccgc ctggagcagc tgcatggcga gctgcgggag
     1861 ctgcacgccc gaatcctgct gcccggccct gggcctggcc cagctgagcc tgtggcctca
     1921 ggaccccggc cgtgggcaga gcagctcagg gctcggcacc tagaggctct ccggaggcag
     1981 ctgcatgtgg agctgaaggt gaagcagggg gctgagaaca tgacccacac gtgcgccagt
     2041 ggcaccccca aggagaggaa gctcctggca gctgcccagc agatgctgcg ggacagccag
     2101 ctgaaggtgg ccctgctgcg gatgaagatc agcagcctgg aggccagtgg gtccccggag
     2161 ccagggcctg agctgctggc ggaggagcta cagcatcgac tgcacgttga ggcagctgtg
     2221 gctgagggcg ccaagaacgt ggtgaaactg cttagtagcc ggagaacaca ggaccgcaag
     2281 gcactggctg aggcccaggc ccagctacag gagtcctctc agaaactgga cctcctgcgc
     2341 ctggccttgg agcagctgct ggagcaactg cctcctgccc accctttgcg cagcagagtg
     2401 acccgagagt tgcgggctgc ggtgcctgga tacccccagc cttcagggac acctgtgaag
     2461 cccaccgccc taacagggac actgcaggtc cgcctcctgg gctgtgaaca gttgctgaca
     2521 gccgtgcctg ggcgctcccc agcggccgca ctggccagca gcccctccga gggctggctt
     2581 cggaccaagg ccaagcacca gcgtggccga ggcgagcttg ccagcgaggt gctggctgtg
     2641 ctaaaggtgg acaaccgtgt tgtggggcag acgggctggg ggcaggtggc cgaacagtcc
     2701 tgggaccaga cctttgtcat cccactggag cgagcccgtg agctggagat tggggtacac
     2761 tggcgggact ggcggcagct atgtggcgtg gccttcctga gacttgagga cttcctggac
     2821 aatgcctgtc accaactgtc cctcagcctg gtaccgcagg gactgctttt tgcccaggtg
     2881 accttctgcg atcctgtcat tgagaggcgg ccccggctgc agaggcagga acgcatcttc
     2941 tctaaacgca gaggccagga cttcctgagg gcttcgcaga tgaacctcgg catggcggcc
     3001 tgggggcgcc tcgtcatgaa cctgctgccc ccctgcagct ccccgagcac aatcagcccc
     3061 cctaaaggat gccctcggac cccaacaaca ctgcgagagg cctctgaccc tgccactccc
     3121 agtaatttcc tgcccaagaa gacccccttg ggtgaagaga tgacaccccc acccaagccc
     3181 ccacgcctct acctccccca ggagccaaca tccgaggaga ctccgcgcac caaacgtccc
     3241 catatggagc ctaggactcg acgtgggcca tctccaccag cctcccccac caggaaaccc
     3301 cctcggcttc aggacttccg ctgcttagct gtgctgggcc ggggacactt tgggaaggtc
     3361 ctcctggtcc agttcaaggg gacagggaaa tactacgcca tcaaagcact gaagaagcag
     3421 gaggtgctca gccgggacga gatagagagc ctgtactgcg agaagcggat cctggaggct
     3481 gtgggctgca cagggcaccc tttcctgctc tccctccttg cctgcttcca gacctccagc
     3541 catgcctgct ttgtgactga gtttgtgcct ggtggtgacc tcatgatgca gatccacgag
     3601 gatgtcttcc ccgagcccca ggcccgcttc tacgtggctt gtgttgtcct ggggctgcag
     3661 ttcttacacg agaagaagat catttacagg gacctgaagt tggataacct tctgctggat
     3721 gcccagggat tcctgaagat cgcagacttt ggactctgca aggaagggat cggcttcggg
     3781 gaccggacta gcaccttctg tggcaccccg gagttcctgg ctcccgaggt gctgacccag
     3841 gaggcataca cacgggctgt ggactggtgg gggctgggtg tgctgctcta cgagatgctg
     3901 gtgggtgagt gcccgttccc aggggacaca gaggaagagg tgtttgactg catcgtcaac
     3961 atggacgccc cctaccccgg ctttctgtcg gtgcaagggc ttgagttcat tcagaagctc
     4021 ctccagaagt gcccggagaa gcgcctcggg gcaggtgagc aggatgccga ggagatcaag
     4081 gtccagccat tcttcaggac caccaactgg caagccctgc tcgcccgcac catccagccc
     4141 cccttcgtgc ctaccctgtg tggccctgcg gacctgcgct actttgaggg cgagttcaca
     4201 gggctgccgc ctgccctgac cccacctgca ccccacagcc tcctcactgc ccgccaacag
     4261 gccgccttcc gggacttcga ctttgtgtca gagcgattcc tggaaccctg aggatcctga
     4321 tatccagcac agtggcggcc gctcgagtct agagggcccg tttaaacccg ctgatcagcc
     4381 tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg
     4441 accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat
     4501 tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag
     4561 gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg
     4621 gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc
     4681 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     4741 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     4801 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     4861 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     4921 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     4981 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
     5041 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt
     5101 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     5161 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
     5221 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     5281 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     5341 tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct
     5401 tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcggat
     5461 ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag
     5521 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg
     5581 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca
     5641 agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc
     5701 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg
     5761 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg
     5821 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta
     5881 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag
     5941 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac
     6001 tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg
     6061 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg
     6121 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg
     6181 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg
     6241 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg
     6301 gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc
     6361 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct
     6421 ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta
     6481 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     6541 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc
     6601 gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     6661 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     6721 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     6781 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     6841 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     6901 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     6961 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     7021 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     7081 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     7141 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     7201 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     7261 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     7321 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     7381 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     7441 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     7501 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     7561 tggtagcggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     7621 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     7681 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     7741 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     7801 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     7861 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     7921 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     7981 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     8041 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     8101 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     8161 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     8221 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     8281 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     8341 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     8401 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     8461 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     8521 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     8581 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     8641 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     8701 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     8761 tccccgaaaa gtgccacctg acgtcgacgg atcgggagat ctcccgatcc cctatggt
//
