LOCUS       V045024                 7920 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V045024
VERSION     V045024
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..3411
                     /label="CHRM3(NM_001347716)"
                     /note="CHRM3(NM_001347716)"
                     /gene="CHRM3"
     polyA_signal    3487..3711
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3757..4185
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4199..4528
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4595..5389
                     /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    5563..5684
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5733..5749)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5757..5773
                     /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(5781..5811)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5826..5847
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6135..6720)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6891..7751)
                     /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(7752..7856)
                     /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 catgaccttg cacaataaca gtacaacctc gcctttgttt
     1681 ccaaacatca gctcctcctg gatacacagc ccctccgatg cagggctgcc cccgggaacc
     1741 gtcactcatt tcggcagcta caatgtttct cgagcagctg gcaatttctc ctctccagac
     1801 ggtaccaccg atgaccctct gggaggtcat accgtctggc aagtggtctt catcgctttc
     1861 ttaacgggca tcctggcctt ggtgaccatc atcggcaaca tcctggtaat tgtgtcattt
     1921 aaggtcaaca agcagctgaa gacggtcaac aactacttcc tcttaagcct ggcctgtgcc
     1981 gatctgatta tcggggtcat ttcaatgaat ctgtttacga cctacatcat catgaatcga
     2041 tgggccttag ggaacttggc ctgtgacctc tggcttgcca ttgactacgt agccagcaat
     2101 gcctctgtta tgaatcttct ggtcatcagc tttgacagat acttttccat cacgaggccg
     2161 ctcacgtacc gagccaaacg aacaacaaag agagccggtg tgatgatcgg tctggcttgg
     2221 gtcatctcct ttgtcctttg ggctcctgcc atcttgttct ggcaatactt tgttggaaag
     2281 agaactgtgc ctccgggaga gtgcttcatt cagttcctca gtgagcccac cattactttt
     2341 ggcacagcca tcgctgcttt ttatatgcct gtcaccatta tgactatttt atactggagg
     2401 atctataagg aaactgaaaa gcgtaccaaa gagcttgctg gcctgcaagc ctctgggaca
     2461 gaggcagaga cagaaaactt tgtccacccc acgggcagtt ctcgaagctg cagcagttac
     2521 gaacttcaac agcaaagcat gaaacgctcc aacaggagga agtatggccg ctgccacttc
     2581 tggttcacaa ccaagagctg gaaacccagc tccgagcaga tggaccaaga ccacagcagc
     2641 agtgacagtt ggaacaacaa tgatgctgct gcctccctgg agaactccgc ctcctccgac
     2701 gaggaggaca ttggctccga gacgagagcc atctactcca tcgtgctcaa gcttccgggt
     2761 cacagcacca tcctcaactc caccaagtta ccctcatcgg acaacctgca ggtgcctgag
     2821 gaggagctgg ggatggtgga cttggagagg aaagccgaca agctgcaggc ccagaagagc
     2881 gtggacgatg gaggcagttt tccaaaaagc ttctccaagc ttcccatcca gctagagtca
     2941 gccgtggaca cagctaagac ttctgacgtc aactcctcag tgggtaagag cacggccact
     3001 ctacctctgt ccttcaagga agccactctg gccaagaggt ttgctctgaa gaccagaagt
     3061 cagatcacta agcggaaaag gatgtccctg gtcaaggaga agaaagcggc ccagaccctc
     3121 agtgcgatct tgcttgcctt catcatcact tggaccccat acaacatcat ggttctggtg
     3181 aacacctttt gtgacagctg catacccaaa accttttgga atctgggcta ctggctgtgc
     3241 tacatcaaca gcaccgtgaa ccccgtgtgc tatgctctgt gcaacaaaac attcagaacc
     3301 actttcaaga tgctgctgct gtgccagtgt gacaaaaaaa agaggcgcaa gcagcagtac
     3361 cagcagagac agtcggtcat ttttcacaag cgcgcacccg agcaggcctt gtaggatcct
     3421 gatatccagc acagtggcgg ccgctcgagt ctagagggcc cgtttaaacc cgctgatcag
     3481 cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct
     3541 tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc
     3601 attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg
     3661 aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg
     3721 cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa
     3781 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
     3841 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     3901 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     3961 aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     4021 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     4081 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct
     4141 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt
     4201 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     4261 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag
     4321 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat
     4381 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     4441 tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg
     4501 cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg
     4561 atctgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc
     4621 aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat
     4681 cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt
     4741 caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg
     4801 gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag
     4861 ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc
     4921 tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc
     4981 tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga
     5041 agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga
     5101 actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg
     5161 cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg
     5221 tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc
     5281 tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc
     5341 cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg
     5401 gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc
     5461 gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc
     5521 ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct
     5581 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     5641 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg
     5701 tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     5761 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     5821 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     5881 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     5941 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     6001 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     6061 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     6121 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     6181 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     6241 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     6301 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     6361 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     6421 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     6481 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     6541 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     6601 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     6661 gctggtagcg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     6721 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     6781 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     6841 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     6901 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     6961 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     7021 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     7081 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     7141 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     7201 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     7261 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     7321 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     7381 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     7441 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     7501 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     7561 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     7621 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     7681 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     7741 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     7801 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     7861 tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atctcccgat cccctatggt
//
