LOCUS       V032807                 8626 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V032807
VERSION     V032807
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        50..429
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        430..633
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             706..3285
                     /label="LDLR(NM_000527)"
                     /note="LDLR(NM_000527)"
                     /gene="LDLR"
     CDS             3292..3318
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             3322..3348
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             3352..3378
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     misc_feature    3402..3990
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4056..4104
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4306..4734
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4748..5077
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5144..5938
                     /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    6114..6235
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6284..6300)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6308..6324
                     /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(6332..6362)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6377..6398
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6686..7274)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7445..8305)
                     /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(8306..8410)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta
       61 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
      121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      421 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      601 cgtgtacggt gggaggtcta tataagcaga gctctggcta gcgtttaaac ttaagcttgg
      661 taccgagctc ggatccacta gtccagtgtg gtggaattcg ccaccatggg gccctggggc
      721 tggaaattgc gctggaccgt cgccttgctc ctcgccgcgg cggggactgc agtgggcgac
      781 agatgcgaaa gaaacgagtt ccagtgccaa gacgggaaat gcatctccta caagtgggtc
      841 tgcgatggca gcgctgagtg ccaggatggc tctgatgagt cccaggagac gtgcttgtct
      901 gtcacctgca aatccgggga cttcagctgt gggggccgtg tcaaccgctg cattcctcag
      961 ttctggaggt gcgatggcca agtggactgc gacaacggct cagacgagca aggctgtccc
     1021 cccaagacgt gctcccagga cgagtttcgc tgccacgatg ggaagtgcat ctctcggcag
     1081 ttcgtctgtg actcagaccg ggactgcttg gacggctcag acgaggcctc ctgcccggtg
     1141 ctcacctgtg gtcccgccag cttccagtgc aacagctcca cctgcatccc ccagctgtgg
     1201 gcctgcgaca acgaccccga ctgcgaagat ggctcggatg agtggccgca gcgctgtagg
     1261 ggtctttacg tgttccaagg ggacagtagc ccctgctcgg ccttcgagtt ccactgccta
     1321 agtggcgagt gcatccactc cagctggcgc tgtgatggtg gccccgactg caaggacaaa
     1381 tctgacgagg aaaactgcgc tgtggccacc tgtcgccctg acgaattcca gtgctctgat
     1441 ggaaactgca tccatggcag ccggcagtgt gaccgggaat atgactgcaa ggacatgagc
     1501 gatgaagttg gctgcgttaa tgtgacactc tgcgagggac ccaacaagtt caagtgtcac
     1561 agcggcgaat gcatcaccct ggacaaagtc tgcaacatgg ctagagactg ccgggactgg
     1621 tcagatgaac ccatcaaaga gtgcgggacc aacgaatgct tggacaacaa cggcggctgt
     1681 tcccacgtct gcaatgacct taagatcggc tacgagtgcc tgtgccccga cggcttccag
     1741 ctggtggccc agcgaagatg cgaagatatc gatgagtgtc aggatcccga cacctgcagc
     1801 cagctctgcg tgaacctgga gggtggctac aagtgccagt gtgaggaagg cttccagctg
     1861 gacccccaca cgaaggcctg caaggctgtg ggctccatcg cctacctctt cttcaccaac
     1921 cggcacgagg tcaggaagat gacgctggac cggagcgagt acaccagcct catccccaac
     1981 ctgaggaacg tggtcgctct ggacacggag gtggccagca atagaatcta ctggtctgac
     2041 ctgtcccaga gaatgatctg cagcacccag cttgacagag cccacggcgt ctcttcctat
     2101 gacaccgtca tcagcagaga catccaggcc cccgacgggc tggctgtgga ctggatccac
     2161 agcaacatct actggaccga ctctgtcctg ggcactgtct ctgttgcgga taccaagggc
     2221 gtgaagagga aaacgttatt cagggagaac ggctccaagc caagggccat cgtggtggat
     2281 cctgttcatg gcttcatgta ctggactgac tggggaactc ccgccaagat caagaaaggg
     2341 ggcctgaatg gtgtggacat ctactcgctg gtgactgaaa acattcagtg gcccaatggc
     2401 atcaccctag atctcctcag tggccgcctc tactgggttg actccaaact tcactccatc
     2461 tcaagcatcg atgtcaacgg gggcaaccgg aagaccatct tggaggatga aaagaggctg
     2521 gcccacccct tctccttggc cgtctttgag gacaaagtat tttggacaga tatcatcaac
     2581 gaagccattt tcagtgccaa ccgcctcaca ggttccgatg tcaacttgtt ggctgaaaac
     2641 ctactgtccc cagaggatat ggttctcttc cacaacctca cccagccaag aggagtgaac
     2701 tggtgtgaga ggaccaccct gagcaatggc ggctgccagt atctgtgcct ccctgccccg
     2761 cagatcaacc cccactcgcc caagtttacc tgcgcctgcc cggacggcat gctgctggcc
     2821 agggacatga ggagctgcct cacagaggct gaggctgcag tggccaccca ggagacatcc
     2881 accgtcaggc taaaggtcag ctccacagcc gtaaggacac agcacacaac cacccgacct
     2941 gttcccgaca cctcccggct gcctggggcc acccctgggc tcaccacggt ggagatagtg
     3001 acaatgtctc accaagctct gggcgacgtt gctggcagag gaaatgagaa gaagcccagt
     3061 agcgtgaggg ctctgtccat tgtcctcccc atcgtgctcc tcgtcttcct ttgcctgggg
     3121 gtcttccttc tatggaagaa ctggcggctt aagaacatca acagcatcaa ctttgacaac
     3181 cccgtctatc agaagaccac agaggatgag gtccacattt gccacaacca ggacggctac
     3241 agctacccct cgagacagat ggtcagtctg gaggatgacg tggcgctcga gtacccatac
     3301 gacgtacctg attatgcaac ttatccttat gatgtacctg attatgctag ctatccttat
     3361 gatgtcccag actacgcata gtgagggccc gatatctcga caatcaacct ctggattaca
     3421 aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat
     3481 acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct
     3541 ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac
     3601 gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca
     3661 cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca
     3721 tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg
     3781 tggtgttgtc ggggaagctg acgtcctttc catggctgct cgcctgtgtt gccacctgga
     3841 ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt
     3901 cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga
     3961 gtcggatctc cctttgggcc gcctccccgc ctggaaacgg gggaggctaa ctgaaacacg
     4021 gaaggagaca ataccggaag gaacccgcgc tatgacggca ataaaaagac agaataaaac
     4081 gcacgggtgt tgggtcgttt gttcataaac gcggggttcg gtcccagggc tggcactctg
     4141 tcgatacccc accgagaccc cattggggcc aatacgcccg cgtttcttcc ttttccccac
     4201 cccacccccc aagttcgggt gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc
     4261 ctgccatagc agatctgcgc agctggggct ctagggggta tccccacgcg ccctgtagcg
     4321 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     4381 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     4441 cccgtcaagc tctaaatcgg ggcatccctt tagggttccg atttagtgct ttacggcacc
     4501 tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga
     4561 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     4621 ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg attttgggga
     4681 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct
     4741 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     4801 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc
     4861 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac
     4921 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     4981 aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta
     5041 gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc
     5101 cattttcgga tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg
     5161 attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca
     5221 acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt
     5281 tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg
     5341 gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga
     5401 agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca
     5461 ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct
     5521 tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac
     5581 tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc
     5641 gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt
     5701 gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt
     5761 catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg
     5821 tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat
     5881 cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc
     5941 gggactctgg ggttcgcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt
     6001 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
     6061 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt
     6121 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     6181 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     6241 tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg
     6301 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     6361 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     6421 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     6481 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     6541 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     6601 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     6661 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     6721 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     6781 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     6841 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca atgctcacgc tgtaggtatc
     6901 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     6961 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     7021 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     7081 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta
     7141 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     7201 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     7261 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     7321 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     7381 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     7441 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     7501 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     7561 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     7621 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     7681 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     7741 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     7801 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     7861 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     7921 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     7981 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     8041 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     8101 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     8161 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     8221 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     8281 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     8341 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     8401 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc
     8461 cgatccccta tggtcgactc tcagtacaat ctgctctgat gccgcatagt taagccagta
     8521 tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac
     8581 aacaaggcaa ggcttgaccg acaattgcat gaagaatctg cttagg
//
