LOCUS       V033041                 7960 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V033041
VERSION     V033041
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..3450
                     /label="GPC3(NM_001164617)"
                     /note="GPC3(NM_001164617)"
                     /gene="GPC3"
     polyA_signal    3527..3751
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3797..4225
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4239..4568
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4635..5429
                     /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    5603..5724
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5773..5789)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5797..5813
                     /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(5821..5851)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5866..5887
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6175..6760)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6931..7791)
                     /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(7792..7896)
                     /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 catggccggg accgtgcgca ccgcgtgctt ggtggtggcg
     1681 atgctgctca gcttggactt cccgggacag gcgcagcccc cgccgccgcc gccggacgcc
     1741 acctgtcacc aagtccgctc cttcttccag agactgcagc ccggactcaa gtgggtgcca
     1801 gaaactcccg tgccaggatc agatttgcaa gtatgtctcc ctaagggccc aacatgctgc
     1861 tcaagaaaga tggaagaaaa ataccaacta acagcacgat tgaacatgga acagctgctt
     1921 cagtctgcaa gtatggagct caagttctta attattcaga atgctgcggt tttccaagag
     1981 gcctttgaaa ttgttgttcg ccatgccaag aactacacca atgccatgtt caagaacaac
     2041 tacccaagcc tgactccaca agcttttgag tttgtgggtg aatttttcac agatgtgtct
     2101 ctctacatct tgggttctga catcaatgta gatgacatgg tcaatgaatt gtttgacagc
     2161 ctgtttccag tcatctatac ccagctaatg aacccaggcc tgcctgattc agccttggac
     2221 atcaatgagt gcctccgagg agcaagacgt gacctgaaag tatttgggaa tttccccaag
     2281 cttattatga cccaggtttc caagtcactg caagtcacta ggatcttcct tcaggctctg
     2341 aatcttggaa ttgaagtgat caacacaact gatcacctga agttcagtaa ggactgtggc
     2401 cgaatgctca ccagaatgtg gtactgctct tactgccagg gactgatgat ggttaaaccc
     2461 tgtggcggtt actgcaatgt ggtcatgcaa ggctgtatgg caggtgtggt ggagattgac
     2521 aagtactgga gagaatacat tctgtccctt gaagaacttg tgaatggcat gtacagaatc
     2581 tatgacatgg agaacgtact gcttggtctc ttttcaacaa tccatgattc tatccagtat
     2641 gtccagaaga atgcaggaaa gctgaccacc actgaaactg agaagaaaat atggcacttc
     2701 aaatatccta tcttcttcct gtgtataggg ctagacttac agattggcaa gttatgtgcc
     2761 cattctcaac aacgccaata tagatctgct tattatcctg aagatctctt tattgacaag
     2821 aaagtattaa aagttgctca tgtagaacat gaagaaacct tatccagccg aagaagggaa
     2881 ctaattcaga agttgaagtc tttcatcagc ttctatagtg ctttgcctgg ctacatctgc
     2941 agccatagcc ctgtggcgga aaacgacacc ctttgctgga atggacaaga actcgtggag
     3001 agatacagcc aaaaggcagc aaggaatgga atgaaaaacc agttcaatct ccatgagctg
     3061 aaaatgaagg gccctgagcc agtggtcagt caaattattg acaaactgaa gcacattaac
     3121 cagctcctga gaaccatgtc tatgcccaaa ggtagagttc tggataaaaa cctggatgag
     3181 gaagggtttg aaagtggaga ctgcggtgat gatgaagatg agtgcattgg aggctctggt
     3241 gatggaatga taaaagtgaa gaatcagctc cgcttccttg cagaactggc ctatgatctg
     3301 gatgtggatg atgcgcctgg aaacagtcag caggcaactc cgaaggacaa cgagataagc
     3361 acctttcaca acctcgggaa cgttcattcc ccgctgaagc ttctcaccag catggccatc
     3421 tcggtggtgt gcttcttctt cctggtgcac tgaggatcct gatatccagc acagtggcgg
     3481 ccgctcgagt ctagagggcc cgtttaaacc cgctgatcag cctcgactgt gccttctagt
     3541 tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact
     3601 cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat
     3661 tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc
     3721 aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc
     3781 tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     3841 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     3901 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     3961 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     4021 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     4081 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     4141 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     4201 atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa
     4261 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     4321 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     4381 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     4441 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     4501 ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     4561 tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa gagacaggat
     4621 gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg
     4681 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg
     4741 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg
     4801 ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc
     4861 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg
     4921 aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca
     4981 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc
     5041 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg
     5101 atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg
     5161 cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata
     5221 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg
     5281 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat
     5341 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct
     5401 tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca
     5461 agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt
     5521 gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat
     5581 gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag
     5641 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     5701 gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt
     5761 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     5821 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     5881 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     5941 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     6001 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     6061 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     6121 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     6181 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     6241 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     6301 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     6361 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     6421 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     6481 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     6541 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     6601 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     6661 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtttttttgt
     6721 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     6781 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     6841 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     6901 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     6961 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     7021 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     7081 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     7141 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     7201 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     7261 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     7321 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     7381 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     7441 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     7501 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     7561 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     7621 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     7681 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     7741 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     7801 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     7861 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     7921 tgacgtcgac ggatcgggag atctcccgat cccctatggt
//
