LOCUS       V031414                 8696 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V031414
VERSION     V031414
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             896..3496
                     /label="E2F8(NM_024680)"
                     /note="E2F8(NM_024680)"
                     /gene="E2F8"
     CDS             3503..4219
                     /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"
     polyA_signal    4263..4487
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4533..4961
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4975..5304
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5371..6165
                     /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    6339..6460
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6509..6525)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6533..6549
                     /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(6557..6587)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6602..6623
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6911..7496)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7667..8527)
                     /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(8528..8632)
                     /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 ggctagcgtt taaacttaag cttggtaccg ccaccatgga
      901 gaacgaaaag gaaaatctct tttgtgagcc acataaaagg ggactaatga aaacacctct
      961 gaaagaatcc accacagcaa atatcgtgtt ggcagagatc cagcctgact ttggcccttt
     1021 aaccacacct accaagccca aggaaggctc tcagggagag ccgtggacac cgacagccaa
     1081 cctgaaaatg ctcatcagtg ctgtgagccc tgagatccgc aacagagatc agaaaagggg
     1141 tttgtttgac aacagaagtg gattacctga ggccaaagac tgtatacacg aacacttatc
     1201 tggagatgaa tttgagaaat cccaaccaag tcgaaaagag aaaagtttag gattattgtg
     1261 tcataagttc ttagcacgat atcctaatta tcccaaccct gctgtgaata atgacatctg
     1321 ccttgacgaa gtggcagagg aacttaatgt tgaacgtcga cgcatttacg atatcgtgaa
     1381 cgtcctagag agtttacata tggtgagccg cctcgccaaa aacaggtaca cttggcacgg
     1441 gcgacacaat ctcaacaaaa cccttggcac cttgaagagc atcggggagg agaataagta
     1501 cgccgagcag attatgatga tcaaaaagaa agaatatgag caagagtttg actttattaa
     1561 gagttacagt atagaggatc atatcatcaa atcaaacact ggcccaaatg gacacccaga
     1621 catgtgtttt gtggaactcc ctggagtgga atttcgggca gcttctgtaa acagccgcaa
     1681 agacaagtct ttaagggtaa tgagccagaa atttgtgatg ctgtttttgg tgtcaacgcc
     1741 tcagatagta agcctagaag ttgctgccaa gattttaatt ggggaggacc atgtggaaga
     1801 tttggataaa agcaagttta aaacaaaaat taggaggttg tatgatatag ctaatgttct
     1861 gagtagcctg gatcttatca agaaagttca tgttacagag gaaagaggcc gaaaaccagc
     1921 tttcaaatgg accggcccag aaatcagtcc aaataccagt ggctccagcc cagtcattca
     1981 ttttactccc tctgatttgg aggtgagacg gtcttcaaaa gagaactgtg ccaaaaacct
     2041 cttttccaca cgtgggaaac caaactttac tcgacaccca tctcttatca aattggtaaa
     2101 gagtatagaa agtgatcgga gaaagataaa ttctgcgccc agtagcccta tcaagaccaa
     2161 caaagctgag agttctcaga attctgcacc cttcccaagt aaaatggctc agctcgcagc
     2221 tatttgtaaa atgcagttag aagagcaatc aagtgaatcc agacagaaag tgaaagtaca
     2281 gctggcaaga tctggaccct gcaaaccagt agcccctctg gaccccccag tgaatgctga
     2341 gatggagctg acagcaccgt ccctcatcca gcccctggga atggttcccc tgatccccag
     2401 ccccttgtca tcagcagtgc ccctgatcct acctcaggcc ccttcaggcc catcctatgc
     2461 catctacctg cagcccactc aagcccacca aagtgtgacg ccaccccaag gcctgagccc
     2521 aacggtgtgc accacccact cttctaaagc tactggctca aaagactcca cagatgccac
     2581 cactgagaag gcagccaatg atacctcaaa ggccagtgcc tctaccaggc ctggaagctt
     2641 gctgccagca ccagagaggc aaggggcaaa gagccgaacc agggagccag ctggagaaag
     2701 aggctcaaag agggcaagca tgctcgagga cagtggttcc aaaaagaaat ttaaagagga
     2761 cctaaaagga cttgaaaatg tctccgcaac cttgttccca tcaggatacc taatccctct
     2821 cacgcagtgc tcatccctgg gggcagagtc cattttgtct ggtaaagaaa actcaagtgc
     2881 tctttcccca aaccacagga tttacagctc cccaattgca ggtgttattc cagtgacatc
     2941 atctgaactc actgctgtta attttccctc ttttcatgta acaccgttga agctaatggt
     3001 ctcaccaact tccgtggcag ccgtacctgt cgggaacagc ccggctctcg cttcaagcca
     3061 ccctgttccc atccagaacc caagctcagc cattgtaaac ttcaccctgc agcacctggg
     3121 actcatctca cccaatgtgc agttgtctgc cagccctggg tctggaatcg ttcctgtgtc
     3181 tccaagaata gagtctgtta atgtcgcacc agaaaatgca ggcactcagc aaggaagggc
     3241 caccaactat gactcaccag tcccaggcca gagccagcca aatggacaat cagttgctgt
     3301 gacaggggca caacagcctg ttcctgtgac acccaaaggg tcacaattag tggccgaaag
     3361 tttcttccgt accccaggtg gacccaccaa gccaaccagc tcatcctgca tggattttga
     3421 gggtgctaat aaaacctcct taggaactct ctttgtccca cagcgaaaac tggaagtctc
     3481 aacagaggat gtccatggat ccatggtgag caagggcgag gagctgttca ccggggtggt
     3541 gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga
     3601 gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa
     3661 gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag
     3721 ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta
     3781 cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt
     3841 gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga
     3901 ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat
     3961 catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga
     4021 ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc
     4081 cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa
     4141 cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg
     4201 catggacgag ctgtacaagc tcgagtctag agggcccgtt taaacccgct gatcagcctc
     4261 gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac
     4321 cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg
     4381 tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga
     4441 ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga
     4501 aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc
     4561 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     4621 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     4681 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     4741 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     4801 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     4861 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg
     4921 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt
     4981 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     5041 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     5101 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     5161 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     5221 tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt
     5281 tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct
     5341 gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt
     5401 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc
     5461 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag
     5521 accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg
     5581 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac
     5641 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc
     5701 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc
     5761 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc
     5821 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg
     5881 ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat
     5941 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc
     6001 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa
     6061 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat
     6121 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt
     6181 tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg
     6241 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc
     6301 agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata
     6361 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     6421 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga
     6481 cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     6541 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     6601 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     6661 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     6721 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     6781 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     6841 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     6901 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     6961 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     7021 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     7081 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     7141 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     7201 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     7261 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     7321 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     7381 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     7441 gtagcggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     7501 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     7561 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     7621 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     7681 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     7741 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     7801 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     7861 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     7921 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     7981 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     8041 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     8101 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     8161 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     8221 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     8281 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     8341 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     8401 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     8461 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     8521 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     8581 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     8641 cccgaaaagt gccacctgac gtcgacggat cgggagatct cccgatcccc tatggt
//
