LOCUS       V030453                 7468 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V030453
VERSION     V030453
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..2958
                     /label="FGL2(NM_006682)"
                     /note="FGL2(NM_006682)"
                     /gene="FGL2"
     polyA_signal    3035..3259
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3305..3733
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3747..4076
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4143..4937
                     /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    5111..5232
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5281..5297)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5305..5321
                     /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(5329..5359)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5374..5395
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5683..6268)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6439..7299)
                     /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(7300..7404)
                     /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 catgaagctg gctaactggt actggctgag ctcagctgtt
     1681 cttgccactt acggtttttt ggttgtggca aacaatgaaa cagaggaaat taaagatgaa
     1741 agagcaaagg atgtctgccc agtgagacta gaaagcagag ggaaatgcga agaggcaggg
     1801 gagtgcccct accaggtaag cctgcccccc ttgactattc agctcccgaa gcaattcagc
     1861 aggatcgagg aggtgttcaa agaagtccaa aacctcaagg aaatcgtaaa tagtctaaag
     1921 aaatcttgcc aagactgcaa gctgcaggct gatgacaacg gagacccagg cagaaacgga
     1981 ctgttgttac ccagtacagg agccccggga gaggttggtg ataacagagt tagagaatta
     2041 gagagtgagg ttaacaagct gtcctctgag ctaaagaatg ccaaagagga gatcaatgta
     2101 cttcatggtc gcctggagaa gctgaatctt gtaaatatga acaacataga aaattatgtt
     2161 gacagcaaag tggcaaatct aacatttgtt gtcaatagtt tggatggcaa atgttcaaag
     2221 tgtcccagcc aagaacaaat acagtcacgt ccagttcaac atctaatata taaagattgc
     2281 tctgactact acgcaatagg caaaagaagc agtgagacct acagagttac acctgatccc
     2341 aaaaatagta gctttgaagt ttactgtgac atggagacca tggggggagg ctggacagtg
     2401 ctgcaggcac gtctcgatgg gagcaccaac ttcaccagaa catggcaaga ctacaaagca
     2461 ggctttggaa acctcagaag ggaattttgg ctggggaacg ataaaattca tcttctgacc
     2521 aagagtaagg aaatgattct gagaatagat cttgaagact ttaatggtgt cgaactatat
     2581 gccttgtatg atcagtttta tgtggctaat gagtttctca aatatcgttt acacgttggt
     2641 aactataatg gcacagctgg agatgcatta cgtttcaaca aacattacaa ccacgatctg
     2701 aagtttttca ccactccaga taaagacaat gatcgatatc cttctgggaa ctgtgggctg
     2761 tactacagtt caggctggtg gtttgatgca tgtctttctg caaacttaaa tggcaaatat
     2821 tatcaccaaa aatacagagg tgtccgtaat gggattttct ggggtacctg gcctggtgta
     2881 agtgaggcac accctggtgg ctacaagtcc tccttcaaag aggctaagat gatgatcaga
     2941 cccaagcact ttaagccata aggatcctga tatccagcac agtggcggcc gctcgagtct
     3001 agagggcccg tttaaacccg ctgatcagcc tcgactgtgc cttctagttg ccagccatct
     3061 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt
     3121 tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg
     3181 ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg
     3241 gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat
     3301 ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     3361 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     3421 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     3481 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     3541 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     3601 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     3661 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     3721 tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
     3781 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa
     3841 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     3901 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     3961 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct
     4021 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc
     4081 tcccgggagc ttgtatatcc attttcggat ctgatcaaga gacaggatga ggatcgtttc
     4141 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
     4201 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
     4261 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
     4321 tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
     4381 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
     4441 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
     4501 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
     4561 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
     4621 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg
     4681 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
     4741 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
     4801 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
     4861 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     4921 ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa
     4981 cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
     5041 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
     5101 cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     5161 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     5221 caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg
     5281 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     5341 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     5401 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     5461 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     5521 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     5581 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     5641 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     5701 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     5761 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     5821 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     5881 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     5941 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     6001 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     6061 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     6121 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     6181 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ttttttgttt gcaagcagca
     6241 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     6301 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     6361 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     6421 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     6481 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     6541 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     6601 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     6661 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     6721 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     6781 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     6841 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     6901 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     6961 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     7021 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     7081 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     7141 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     7201 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     7261 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     7321 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     7381 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg
     7441 atcgggagat ctcccgatcc cctatggt
//
