LOCUS       V032311                 8629 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V032311
VERSION     V032311
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..3381
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     misc_feature    3405..3993
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4059..4107
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4309..4737
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4751..5080
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5147..5941
                     /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    6117..6238
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6287..6303)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6311..6327
                     /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(6335..6365)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6380..6401
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6689..7277)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7448..8308)
                     /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(8309..8413)
                     /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 ggaacaaaaa
     3301 ctaatatcag aggaagattt ggaacagaaa ctaatcagtg aagaagattt agagcaaaag
     3361 ttaatttccg aggaggactt atagtgaggg cccgatatct cgacaatcaa cctctggatt
     3421 acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg
     3481 gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct
     3541 cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc
     3601 aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca
     3661 ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac
     3721 tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt
     3781 ccgtggtgtt gtcggggaag ctgacgtcct ttccatggct gctcgcctgt gttgccacct
     3841 ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc
     3901 cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga
     3961 cgagtcggat ctccctttgg gccgcctccc cgcctggaaa cgggggaggc taactgaaac
     4021 acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa
     4081 aacgcacggg tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact
     4141 ctgtcgatac cccaccgaga ccccattggg gccaatacgc ccgcgtttct tccttttccc
     4201 caccccaccc cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag
     4261 gccctgccat agcagatctg cgcagctggg gctctagggg gtatccccac gcgccctgta
     4321 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     4381 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     4441 ttccccgtca agctctaaat cggggcatcc ctttagggtt ccgatttagt gctttacggc
     4501 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
     4561 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     4621 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgg
     4681 ggatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat
     4741 tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag
     4801 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc
     4861 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct
     4921 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     4981 actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa
     5041 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat
     5101 atccattttc ggatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga
     5161 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc
     5221 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc
     5281 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc
     5341 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac
     5401 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc
     5461 tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac
     5521 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg
     5581 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct
     5641 cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt
     5701 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg
     5761 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac
     5821 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg
     5881 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg
     5941 agcgggactc tggggttcgc gaaatgaccg accaagcgac gcccaacctg ccatcacgag
     6001 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg
     6061 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact
     6121 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     6181 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     6241 atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc
     6301 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     6361 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     6421 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     6481 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     6541 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     6601 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     6661 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     6721 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     6781 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     6841 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcaatgctca cgctgtaggt
     6901 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     6961 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     7021 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     7081 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg
     7141 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     7201 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     7261 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     7321 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     7381 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     7441 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     7501 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     7561 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     7621 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     7681 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     7741 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     7801 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     7861 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     7921 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     7981 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     8041 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     8101 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     8161 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     8221 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     8281 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     8341 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     8401 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc
     8461 tcccgatccc ctatggtcga ctctcagtac aatctgctct gatgccgcat agttaagcca
     8521 gtatctgctc cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca aaatttaagc
     8581 tacaacaagg caaggcttga ccgacaattg catgaagaat ctgcttagg
//
