LOCUS       V024162                10292 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V024162
VERSION     V024162
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        235..614
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        615..818
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        863..881
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             941..5782
                     /label="Lrp5(NM_008513)"
                     /note="Lrp5(NM_008513)"
                     /gene="Lrp5"
     CDS             5789..5854
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    5892..6116
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      6162..6590
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        6604..6933
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             7000..7794
                     /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    7968..8089
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(8138..8154)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    8162..8178
                     /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(8186..8216)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    8231..8252
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(8540..9125)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(9296..10156)
                     /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(10157..10261)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttggta ccgagctcgg atccgccacc atggaaacgg cgccgacccg
      961 ggcccctccg ccgccgccgc cgccgctgct gctgctggtg ctgtactgca gcttggtccc
     1021 cgccgcggcc tcaccgctcc tgttgtttgc caaccgccgg gatgtgcggc tagtggatgc
     1081 cggcggagtg aagctggagt ccaccattgt ggccagtggc ctggaggatg cagctgctgt
     1141 agacttccag ttctccaagg gtgctgtgta ctggacagat gtgagcgagg aggccatcaa
     1201 acagacctac ctgaaccaga ctggagctgc tgcacagaac attgtcatct cgggcctcgt
     1261 gtcacctgat ggcctggcct gtgactgggt tggcaagaag ctgtactgga cggactccga
     1321 gaccaaccgc attgaggttg ccaacctcaa tgggacgtcc cgtaaggttc tcttctggca
     1381 ggacctggac cagccaaggg ccattgccct ggatcctgca catgggtaca tgtactggac
     1441 tgactggggg gaagcacccc ggatcgagcg ggcagggatg gatggcagta cccggaagat
     1501 cattgtagac tccgacattt actggcccaa tgggctgacc atcgacctgg aggaacagaa
     1561 gctgtactgg gccgatgcca agctcagctt catccaccgt gccaacctgg acggctcctt
     1621 ccggcagaag gtggtggagg gcagcctcac tcaccctttt gccctgacac tctctgggga
     1681 cacactctac tggacagact ggcagacccg ctccatccac gcctgcaaca agtggacagg
     1741 ggagcagagg aaggagatcc ttagtgctct gtactcaccc atggacatcc aagtgctgag
     1801 ccaggagcgg cagcctccct tccacacacc atgcgaggag gacaacggtg gctgttccca
     1861 cctgtgcctg ctgtccccga gggagccttt ctactcctgt gcctgcccca ctggtgtgca
     1921 gttgcaggac aatggcaaga cgtgcaagac aggggctgag gaagtgctgc tgctggctcg
     1981 gaggacagac ctgaggagga tctctctgga cacccctgac ttcacagaca tagtgctgca
     2041 ggtgggcgac atccggcatg ccattgccat tgactacgat cccctggagg gctacgtgta
     2101 ctggaccgac gatgaggtgc gggctatccg cagggcgtac ctagatggct caggtgcgca
     2161 gacacttgtg aacactgaga tcaatgaccc cgatggcatt gctgtggact gggtcgcccg
     2221 gaacctctac tggacagata caggcactga cagaattgag gtgactcgcc tcaacggcac
     2281 ctcccgaaag atcctggtat ctgaggacct ggacgaaccg cgagccattg tgttgcaccc
     2341 tgtgatgggc ctcatgtact ggacagactg gggggagaac cccaaaatcg aatgcgccaa
     2401 cctagatggg agagatcggc atgtcctggt gaacacctcc cttgggtggc ccaacggact
     2461 ggccctggac ctgcaggagg gcaagctgta ctggggggat gccaaaactg ataaaatcga
     2521 ggtgatcaac atagatggga caaagcggaa gaccctgctt gaggacaagc tcccacatat
     2581 ttttgggttc acactgctgg gggacttcat ctactggacc gactggcaga gacgcagtat
     2641 tgaaagggtc cacaaggtca aggccagccg ggatgtcatc attgatcaac tccccgacct
     2701 gatgggactc aaagctgtga atgtggccaa ggttgtcgga accaacccat gtgcggatgg
     2761 aaatggaggg tgcagccatc tgtgcttctt caccccacgt gccaccaagt gtggctgccc
     2821 cattggcctg gagctgttga gtgacatgaa gacctgcata atccccgagg ccttcctggt
     2881 attcaccagc agagccacca tccacaggat ctccctggag actaacaaca acgatgtggc
     2941 tatcccactc acgggtgtca aagaggcctc tgcactggac tttgatgtgt ccaacaatca
     3001 catctactgg actgatgtca gcctcaagac gatcagccga gccttcatga atgggagctc
     3061 agtggagcac gtgattgagt ttggcctcga ctaccctgaa ggaatggctg tggactggat
     3121 gggcaagaac ctctattggg cggacacagg gaccaacagg attgaggtgg cccggctgga
     3181 tgggcagttc cggcaggtgc ttgtgtggag agaccttgac aaccccaggt ctctggctct
     3241 ggatcctact aaaggctaca tctactggac tgagtggggt ggcaagccaa ggattgtgcg
     3301 ggccttcatg gatgggacca attgtatgac actggtagac aaggtgggcc gggccaacga
     3361 cctcaccatt gattatgccg accagcgact gtactggact gacctggaca ccaacatgat
     3421 tgagtcttcc aacatgctgg gtcaggagcg catggtgata gctgacgatc tgccctaccc
     3481 gtttggcctg actcaatata gcgattacat ctactggact gactggaacc tgcatagcat
     3541 tgaacgggcg gacaagacca gtgggcggaa ccgcaccctc atccagggtc acctggactt
     3601 cgtcatggac atcctggtgt tccactcctc ccgtcaggat ggcctcaacg actgcgtgca
     3661 cagcaatggc cagtgtgggc agctgtgcct cgccatcccc ggaggccacc gctgtggctg
     3721 tgcttcacac tacacgctgg accccagcag ccgcaactgc agcccgccct ccaccttctt
     3781 gctgttcagc cagaaatttg ccatcagccg gatgatcccc gatgaccagc tcagcccgga
     3841 ccttgtccta ccccttcatg ggctgaggaa cgtcaaagcc atcaactatg acccgctgga
     3901 caagttcatc tactgggtgg acgggcgcca gaacatcaag agggccaagg acgacggtac
     3961 ccagccctcc atgctgacct ctcccagcca aagcctgagc ccagacagac agccacacga
     4021 cctcagcatt gacatctaca gccggacact gttctggacc tgtgaggcca ccaacactat
     4081 caatgtccac cggctggatg gggatgccat gggagtggtg cttcgagggg accgtgacaa
     4141 gccaagggcc attgctgtca atgctgagcg agggtacatg tactttacca acatgcagga
     4201 ccatgctgcc aagatcgagc gagcctccct ggatggcaca gagcgggagg tcctcttcac
     4261 cacaggcctc atccgtcccg tggcccttgt ggtggacaat gctctgggca agctcttctg
     4321 ggtggatgcc gacctaaagc gaatcgaaag ctgtgacctc tctggggcca accgcctgac
     4381 cctggaagat gccaacatcg tacagccagt aggtctgaca gtgctgggca ggcacctcta
     4441 ctggatcgac cgccagcagc agatgatcga gcgcgtggag aagaccactg gggacaagcg
     4501 gactagggtt cagggccgtg tcacccacct gacaggcatc catgccgtgg aggaagtcag
     4561 cctggaggag ttctcagccc atccttgtgc ccgagacaat ggcggctgct cccacatctg
     4621 tatcgccaag ggtgatggaa caccgcgctg ctcgtgccct gtccacctgg tgctcctgca
     4681 gaacctgctg acttgtggtg agcctcctac ctgctcccct gatcagtttg catgtaccac
     4741 tggtgagatc gactgcatcc ccggagcctg gcgctgtgac ggcttccctg agtgtgctga
     4801 ccagagtgat gaagaaggct gcccagtgtg ctccgcctct cagttcccct gcgctcgagg
     4861 ccagtgtgtg gacctgcggt tacgctgcga cggtgaggcc gactgccagg atcgctctga
     4921 tgaagctaac tgcgatgctg tctgtctgcc caatcagttc cggtgcacca gcggccagtg
     4981 tgtcctcatc aagcaacagt gtgactcctt ccccgactgt gctgatgggt ctgatgagct
     5041 catgtgtgaa atcaacaagc caccctctga tgacatccca gcccacagca gtgccattgg
     5101 gcccgtcatt ggtatcatcc tctccctctt cgtcatgggc ggggtctact ttgtctgcca
     5161 gcgtgtgatg tgccagcgct acacaggggc cagtgggccc tttccccacg agtatgttgg
     5221 tggagcccct catgtgcctc tcaacttcat agccccaggt ggctcacagc acggtccctt
     5281 cccaggcatc ccgtgcagca agtccgtgat gagctccatg agcctggtgg gggggcgcgg
     5341 cagtgtgccc ctctatgacc ggaatcacgt cactggggcc tcatccagca gctcgtccag
     5401 cacaaaggcc acactatatc cgccgatcct gaacccaccc ccgtccccgg ccacagaccc
     5461 ctctctctac aacgtggacg tgttttattc ttcaggcatc ccggccaccg ctagaccata
     5521 caggccctac gtcattcgag gtatggcacc cccaacaaca ccgtgcagca cagatgtgtg
     5581 tgacagtgac tacagcacca gtcgctggaa gagcagcaaa tactacctgg acttgaattc
     5641 ggactcagac ccctaccccc ccccgcccac cccccacagc cagtacctat ctgcagagga
     5701 cagctgccca ccctcaccag gcactgagag gagttactgc cacctcttcc cgcccccacc
     5761 gtccccctgc acggactcgt ccctcgagga ctacaaagac catgacggtg attataaaga
     5821 tcatgacatc gactacaagg atgacgatga caagtagtga gggcccgttt aaacccgctg
     5881 atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc
     5941 ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc
     6001 atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa
     6061 gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc
     6121 tgaggcggaa agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc
     6181 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     6241 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     6301 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     6361 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     6421 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     6481 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
     6541 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg
     6601 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     6661 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
     6721 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     6781 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     6841 ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga
     6901 ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt
     6961 ttcggatctg atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg
     7021 cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag
     7081 acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt
     7141 tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta
     7201 tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg
     7261 ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt
     7321 gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat
     7381 ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg
     7441 atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca
     7501 gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc
     7561 catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc
     7621 gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat
     7681 attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc
     7741 gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga
     7801 ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt
     7861 ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga
     7921 tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg
     7981 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     8041 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta
     8101 taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa
     8161 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     8221 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     8281 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     8341 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     8401 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     8461 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     8521 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     8581 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     8641 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     8701 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     8761 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     8821 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     8881 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     8941 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     9001 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     9061 ccaccgctgg tagcggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     9121 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     9181 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     9241 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     9301 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     9361 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     9421 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     9481 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     9541 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     9601 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     9661 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     9721 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     9781 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     9841 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     9901 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     9961 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
    10021 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
    10081 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
    10141 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
    10201 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
    10261 gcacatttcc ccgaaaagtg ccacctgacg tc
//
