LOCUS       V041715                 7738 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V041715
VERSION     V041715
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..3228
                     /label="TYR(NM_000372)"
                     /note="TYR(NM_000372)"
                     /gene="TYR"
     polyA_signal    3305..3529
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3575..4003
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4017..4346
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4413..5207
                     /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    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..6538)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6709..7569)
                     /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(7570..7674)
                     /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 catgctcctg gctgttttgt actgcctgct gtggagtttc
     1681 cagacctccg ctggccattt ccctagagcc tgtgtctcct ctaagaacct gatggagaag
     1741 gaatgctgtc caccgtggag cggggacagg agtccctgtg gccagctttc aggcagaggt
     1801 tcctgtcaga atatccttct gtccaatgca ccacttgggc ctcaatttcc cttcacaggg
     1861 gtggatgacc gggagtcgtg gccttccgtc ttttataata ggacctgcca gtgctctggc
     1921 aacttcatgg gattcaactg tggaaactgc aagtttggct tttggggacc aaactgcaca
     1981 gagagacgac tcttggtgag aagaaacatc ttcgatttga gtgccccaga gaaggacaaa
     2041 ttttttgcct acctcacttt agcaaagcat accatcagct cagactatgt catccccata
     2101 gggacctatg gccaaatgaa aaatggatca acacccatgt ttaacgacat caatatttat
     2161 gacctctttg tctggatgca ttattatgtg tcaatggatg cactgcttgg gggatctgaa
     2221 atctggagag acattgattt tgcccatgaa gcaccagctt ttctgccttg gcatagactc
     2281 ttcttgttgc ggtgggaaca agaaatccag aagctgacag gagatgaaaa cttcactatt
     2341 ccatattggg actggcggga tgcagaaaag tgtgacattt gcacagatga gtacatggga
     2401 ggtcagcacc ccacaaatcc taacttactc agcccagcat cattcttctc ctcttggcag
     2461 attgtctgta gccgattgga ggagtacaac agccatcagt ctttatgcaa tggaacgccc
     2521 gagggacctt tacggcgtaa tcctggaaac catgacaaat ccagaacccc aaggctcccc
     2581 tcttcagctg atgtagaatt ttgcctgagt ttgacccaat atgaatctgg ttccatggat
     2641 aaagctgcca atttcagctt tagaaataca ctggaaggat ttgctagtcc acttactggg
     2701 atagcggatg cctctcaaag cagcatgcac aatgccttgc acatctatat gaatggaaca
     2761 atgtcccagg tacagggatc tgccaacgat cctatcttcc ttcttcacca tgcatttgtt
     2821 gacagtattt ttgagcagtg gctccgaagg caccgtcctc ttcaagaagt ttatccagaa
     2881 gccaatgcac ccattggaca taaccgggaa tcctacatgg ttccttttat accactgtac
     2941 agaaatggtg atttctttat ttcatccaaa gatctgggct atgactatag ctatctacaa
     3001 gattcagacc cagactcttt tcaagactac attaagtcct atttggaaca agcgagtcgg
     3061 atctggtcat ggctccttgg ggcggcgatg gtaggggccg tcctcactgc cctgctggca
     3121 gggcttgtga gcttgctgtg tcgtcacaag agaaagcagc ttcctgaaga aaagcagcca
     3181 ctcctcatgg agaaagagga ttaccacagc ttgtatcaga gccatttata aggatcctga
     3241 tatccagcac agtggcggcc gctcgagtct agagggcccg tttaaacccg ctgatcagcc
     3301 tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg
     3361 accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat
     3421 tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag
     3481 gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg
     3541 gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc
     3601 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     3661 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     3721 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     3781 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     3841 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     3901 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
     3961 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt
     4021 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     4081 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
     4141 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     4201 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     4261 tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct
     4321 tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcggat
     4381 ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag
     4441 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg
     4501 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca
     4561 agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc
     4621 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg
     4681 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg
     4741 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta
     4801 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag
     4861 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac
     4921 tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg
     4981 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg
     5041 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg
     5101 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg
     5161 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg
     5221 gttcgaaatg accgaccaag cgacgcccaa cctgccatca 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 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     6541 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     6601 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     6661 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     6721 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     6781 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     6841 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     6901 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     6961 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     7021 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     7081 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     7141 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     7201 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     7261 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     7321 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     7381 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     7441 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     7501 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     7561 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     7621 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     7681 tccccgaaaa gtgccacctg acgtcgacgg atcgggagat ctcccgatcc cctatggt
//
