LOCUS       V030006                 7963 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V030006
VERSION     V030006
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..3453
                     /label="Ptgs2(NM_011198)"
                     /note="Ptgs2(NM_011198)"
                     /gene="Ptgs2"
     polyA_signal    3530..3754
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3800..4228
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4242..4571
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4638..5432
                     /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    5606..5727
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5776..5792)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5800..5816
                     /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(5824..5854)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5869..5890
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6178..6763)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6934..7794)
                     /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(7795..7899)
                     /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 catgctcttc cgagctgtgc tgctctgcgc tgccctgggg
     1681 ctcagccagg cagcaaatcc ttgctgttcc aatccatgtc aaaaccgtgg ggaatgtatg
     1741 agcacaggat ttgaccagta taagtgtgac tgtacccgga ctggattcta tggtgaaaac
     1801 tgtactacac ctgaatttct gacaagaatc aaattactgc tgaagcccac cccaaacaca
     1861 gtgcactaca tcctgaccca cttcaaggga gtctggaaca ttgtgaacaa catccccttc
     1921 ctgcgaagtt taatcatgaa atatgtgctg acatccagat catatttgat tgacagtcca
     1981 cctacttaca atgtgcacta tggttacaaa agctgggaag ccttctccaa cctctcctac
     2041 tacaccaggg cccttcctcc cgtagcagat gactgcccaa ctcccatggg tgtgaaggga
     2101 aataaggagc ttcctgattc aaaagaagtg ctggaaaagg ttcttctacg gagagagttc
     2161 atccctgacc cccaaggctc aaatatgatg tttgcattct ttgcccagca cttcacccat
     2221 cagtttttca agacagatca taagcgagga cctgggttca cccgaggact gggccatgga
     2281 gtggacttaa atcacattta tggtgaaact ctggacagac aacataaact gcgccttttc
     2341 aaggatggaa aattgaaata tcaggtcatt ggtggagagg tgtatccccc cacagtcaaa
     2401 gacactcagg tagagatgat ctaccctcct cacatccctg agaacctgca gtttgctgtg
     2461 gggcaggaag tctttggtct ggtgcctggt ctgatgatgt atgccaccat ctggcttcgg
     2521 gagcacaaca gagtgtgcga catactcaag caggagcatc ctgagtgggg tgatgagcaa
     2581 ctattccaaa ccagcagact catactcata ggagagacta tcaagatagt gatcgaagac
     2641 tacgtgcaac acctgagcgg ttaccacttc aaactcaagt ttgacccaga gctccttttc
     2701 aaccagcagt tccagtatca gaaccgcatt gcctctgaat tcaacacact ctatcactgg
     2761 caccccctgc tgcccgacac cttcaacatt gaagaccagg agtacagctt caaacagttt
     2821 ctctacaaca actccatcct cctggaacat ggactcactc agtttgttga gtcattcacc
     2881 agacagattg ctggccgggt tgctggggga agaaatgtgc caattgctgt acaagcagtg
     2941 gcaaaggcct ccattgacca gagcagagag atgaaatacc agtctctcaa tgagtaccgc
     3001 aaacgcttct ccctgaagcc gtacacatca tttgaagaac ttacaggaga gaaggaaatg
     3061 gctgcagaat tgaaagccct ctacagtgac atcgatgtca tggaactgta ccctgccctg
     3121 ctggtggaaa aacctcgtcc agatgctatc tttggggaga ccatggtaga gcttggagca
     3181 ccattctcct tgaaaggact tatgggaaat cccatctgtt ctcctcaata ctggaagccg
     3241 agcacctttg gaggcgaagt gggttttaag atcatcaata ctgcctcaat tcagtctctc
     3301 atctgcaata atgtgaaggg gtgtcccttc acttctttca atgtgcaaga tccacagcct
     3361 accaaaacag ccaccatcaa tgcaagtgcc tcccactcca gactagatga cattaaccct
     3421 acagtactaa tcaaaaggcg ttcaactgag ctgtaaggat cctgatatcc agcacagtgg
     3481 cggccgctcg agtctagagg gcccgtttaa acccgctgat cagcctcgac tgtgccttct
     3541 agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc
     3601 actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt
     3661 cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat
     3721 agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg
     3781 ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     3841 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     3901 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     3961 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     4021 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     4081 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     4141 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     4201 ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg
     4261 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     4321 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     4381 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     4441 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     4501 aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     4561 gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat caagagacag
     4621 gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt
     4681 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg
     4741 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg
     4801 gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg
     4861 ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg
     4921 gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca
     4981 tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc
     5041 accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc
     5101 aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca
     5161 aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga
     5221 atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg
     5281 cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg
     5341 aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg
     5401 ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga
     5461 ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag
     5521 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
     5581 catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata
     5641 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     5701 tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag
     5761 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     5821 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     5881 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     5941 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     6001 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     6061 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     6121 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     6181 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     6241 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     6301 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     6361 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     6421 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     6481 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     6541 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     6601 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     6661 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtttttt
     6721 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     6781 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     6841 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     6901 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     6961 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     7021 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     7081 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     7141 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     7201 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     7261 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     7321 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     7381 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     7441 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     7501 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     7561 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     7621 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     7681 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     7741 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     7801 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     7861 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     7921 acctgacgtc gacggatcgg gagatctccc gatcccctat ggt
//
