LOCUS       V033091                 7531 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V033091
VERSION     V033091
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..3021
                     /label="ANGPTL3(NM_014495)"
                     /note="ANGPTL3(NM_014495)"
                     /gene="ANGPTL3"
     polyA_signal    3098..3322
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3368..3796
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3810..4139
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4206..5000
                     /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    5174..5295
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5344..5360)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5368..5384
                     /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(5392..5422)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5437..5458
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5746..6331)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6502..7362)
                     /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(7363..7467)
                     /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 catgttcaca attaagctcc ttctttttat tgttcctcta
     1681 gttatttcct ccagaattga tcaagacaat tcatcatttg attctctatc tccagagcca
     1741 aaatcaagat ttgctatgtt agacgatgta aaaattttag ccaatggcct ccttcagttg
     1801 ggacatggtc ttaaagactt tgtccataag acgaagggcc aaattaatga catatttcaa
     1861 aaactcaaca tatttgatca gtctttttat gatctatcgc tgcaaaccag tgaaatcaaa
     1921 gaagaagaaa aggaactgag aagaactaca tataaactac aagtcaaaaa tgaagaggta
     1981 aagaatatgt cacttgaact caactcaaaa cttgaaagcc tcctagaaga aaaaattcta
     2041 cttcaacaaa aagtgaaata tttagaagag caactaacta acttaattca aaatcaacct
     2101 gaaactccag aacacccaga agtaacttca cttaaaactt ttgtagaaaa acaagataat
     2161 agcatcaaag accttctcca gaccgtggaa gaccaatata aacaattaaa ccaacagcat
     2221 agtcaaataa aagaaataga aaatcagctc agaaggacta gtattcaaga acccacagaa
     2281 atttctctat cttccaagcc aagagcacca agaactactc cctttcttca gttgaatgaa
     2341 ataagaaatg taaaacatga tggcattcct gctgaatgta ccaccattta taacagaggt
     2401 gaacatacaa gtggcatgta tgccatcaga cccagcaact ctcaagtttt tcatgtctac
     2461 tgtgatgtta tatcaggtag tccatggaca ttaattcaac atcgaataga tggatcacaa
     2521 aacttcaatg aaacgtggga gaactacaaa tatggttttg ggaggcttga tggagaattt
     2581 tggttgggcc tagagaagat atactccata gtgaagcaat ctaattatgt tttacgaatt
     2641 gagttggaag actggaaaga caacaaacat tatattgaat attcttttta cttgggaaat
     2701 cacgaaacca actatacgct acatctagtt gcgattactg gcaatgtccc caatgcaatc
     2761 ccggaaaaca aagatttggt gttttctact tgggatcaca aagcaaaagg acacttcaac
     2821 tgtccagagg gttattcagg aggctggtgg tggcatgatg agtgtggaga aaacaaccta
     2881 aatggtaaat ataacaaacc aagagcaaaa tctaagccag agaggagaag aggattatct
     2941 tggaagtctc aaaatggaag gttatactct ataaaatcaa ccaaaatgtt gatccatcca
     3001 acagattcag aaagctttga atgaggatcc tgatatccag cacagtggcg gccgctcgag
     3061 tctagagggc ccgtttaaac ccgctgatca gcctcgactg tgccttctag ttgccagcca
     3121 tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc
     3181 ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg
     3241 gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct
     3301 ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg
     3361 tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     3421 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     3481 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     3541 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
     3601 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     3661 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
     3721 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
     3781 aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
     3841 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
     3901 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     3961 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     4021 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg
     4081 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
     4141 agctcccggg agcttgtata tccattttcg gatctgatca agagacagga tgaggatcgt
     4201 ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc
     4261 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc
     4321 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg
     4381 aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag
     4441 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg
     4501 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg
     4561 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac
     4621 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg
     4681 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc
     4741 ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg
     4801 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc
     4861 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc
     4921 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc
     4981 ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc
     5041 caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg
     5101 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt
     5161 cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     5221 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     5281 catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc
     5341 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     5401 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     5461 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     5521 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     5581 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     5641 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     5701 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     5761 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     5821 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     5881 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     5941 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     6001 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     6061 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     6121 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     6181 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     6241 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtttttttg tttgcaagca
     6301 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     6361 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     6421 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     6481 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     6541 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     6601 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     6661 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     6721 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     6781 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     6841 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     6901 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     6961 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     7021 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     7081 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     7141 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     7201 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     7261 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     7321 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     7381 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     7441 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcga
     7501 cggatcggga gatctcccga tcccctatgg t
//
