LOCUS       V036952                 8309 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V036952
VERSION     V036952
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             907..3759
                     /label="LGR4(NM_018490)"
                     /note="LGR4(NM_018490)"
                     /gene="LGR4"
     CDS             3766..3807
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /transl_table=1
                     /translation="GKPIPNPLLGLDST"
                     /label="V5 tag"
     polyA_signal    3909..4133
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4179..4607
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4621..4950
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5017..5811
                     /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    5985..6106
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6155..6171)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6179..6195
                     /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(6203..6233)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6248..6269
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6557..7142)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7313..8173)
                     /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(8174..8278)
                     /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 gccaccatgc cgggcccgct agggctgctc tgcttcctcg ccctggggct gctcggctcg
      961 gccgggccca gcggcgcggc gccgcctctc tgcgcggcgc cctgcagctg cgacggcgac
     1021 cgtcgggtgg actgctccgg gaaggggctg acggccgtgc ccgaggggct cagcgccttc
     1081 acccaagcgc tggatatcag tatgaacaac attactcagt tgccagaaga tgcatttaag
     1141 aactttcctt ttctagaaga gctacaattg gcgggcaacg acctttcttt tatccaccca
     1201 aaggccttgt ctgggttgaa agaactcaaa gttctaacgc tccagaataa tcagttgaaa
     1261 acagtaccca gtgaagccat tcgagggctg agtgctttgc agtctttgcg tttagatgcc
     1321 aaccatatta cctcagtccc cgaggacagt tttgaaggac ttgttcagtt acggcatctg
     1381 tggctggatg acaacagctt gacggaggtg cctgtgcacc ccctcagcaa tctgcccacc
     1441 ctacaggcgc tgaccctggc tctcaacaag atctcaagca tccctgactt tgcatttacc
     1501 aacctttcaa gcctggtagt tctgcatctt cataacaata aaattagaag cctgagtcaa
     1561 cactgttttg atggactaga taacctggag accttagact tgaattataa taacttgggg
     1621 gaatttcctc aggctattaa agcccttcct agccttaaag agctaggatt tcatagtaat
     1681 tctatttctg ttatccctga tggagcattt gatggtaatc cactcttaag aactatacat
     1741 ttgtatgata atcctctgtc ttttgtgggg aactcagcat ttcacaattt atctgatctt
     1801 cattccctag tcattcgtgg tgcaagcatg gtgcagcagt tccccaatct tacaggaact
     1861 gtccacctgg aaagtctgac tttgacaggt acaaagataa gcagcatacc taataatttg
     1921 tgtcaagaac aaaagatgct taggactttg gacttgtctt acaataatat aagagacctt
     1981 ccaagtttta atggttgcca tgctctggaa gaaatttctt tacagcgtaa tcaaatctac
     2041 caaataaagg aaggcacctt tcaaggcctg atatctctaa ggattctaga tctgagtaga
     2101 aacctgatac atgaaattca cagtagagct tttgccacac ttgggccaat aactaaccta
     2161 gatgtaagtt tcaatgaatt aacttccttt cctacggaag gcctgaatgg gctaaatcaa
     2221 ctgaaacttg tgggcaactt caagctgaaa gaagccttag cagcaaaaga ctttgttaac
     2281 ctcaggtctt tatcagtacc atatgcttat cagtgctgtg cattttgggg ttgtgactct
     2341 tatgcaaatt taaacacaga agataacagc ctccaggacc acagtgtggc acaggagaaa
     2401 ggtactgctg atgcagcaaa tgtcacaagc actcttgaaa atgaagaaca tagtcaaata
     2461 attatccatt gtacaccttc aacaggtgct tttaagccct gtgaatattt actgggaagc
     2521 tggatgattc gtcttactgt gtggttcatt ttcttggttg cattattttt caacctgctt
     2581 gttattttaa caacatttgc atcttgtaca tcactgcctt cgtccaaatt gtttataggc
     2641 ttgatttctg tgtctaactt attcatggga atctatactg gcatcctaac ttttcttgat
     2701 gctgtgtcct ggggcagatt cgctgaattt ggcatttggt gggaaactgg cagtggctgc
     2761 aaagtagctg ggtttcttgc agttttctcc tcagaaagtg ccatattttt attaatgcta
     2821 gcaactgtcg aaagaagctt atctgcaaaa gatataatga aaaatgggaa gagcaatcat
     2881 ctcaaacagt tccgggttgc tgcccttttg gctttcctag gtgctacagt agcaggctgt
     2941 tttccccttt tccatagagg ggaatattct gcatcacccc tttgtttgcc atttcctaca
     3001 ggtgaaacgc catcattagg attcactgta acgttagtgc tattaaactc actagcattt
     3061 ttattaatgg ccgttatcta cactaaacta tactgcaact tggaaaaaga ggacctctca
     3121 gaaaactcac aatctagcat gattaagcat gtcgcttggc taatcttcac caattgcatc
     3181 tttttctgcc ctgtggcgtt tttttcattt gcaccattga tcactgcaat ctctatcagc
     3241 cccgaaataa tgaagtctgt tactctgata ttttttccat tgcctgcttg cctgaatcca
     3301 gtcctgtatg ttttcttcaa cccaaagttt aaagaagact ggaagttact gaagcgacgt
     3361 gttaccaaga aaagtggatc agtttcagtt tccatcagta gccaaggtgg ttgtctggaa
     3421 caggatttct actacgactg tggcatgtac tcacatttgc agggcaacct gactgtttgc
     3481 gactgctgcg aatcgtttct tttaacaaag ccagtatcat gcaaacactt gataaaatca
     3541 cacagctgtc ctgcattggc agtggcttct tgccaaagac ctgagggcta ctggtccgac
     3601 tgtggcacac agtcggccca ctctgattat gcagatgaag aagattcctt tgtctcagac
     3661 agttctgacc aggtgcaggc ctgtggacga gcctgcttct accagagtag aggattccct
     3721 ttggtgcgct atgcttacaa tctaccaaga gttaaagaca agcttggtaa gcctatccct
     3781 aaccctctcc tcggtctcga ttctacatag gatccactag tccagtgtgg tggaattctg
     3841 cagatatcca gcacagtggc ggccgctcga gtctagaggg cccgtttaaa cccgctgatc
     3901 agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc
     3961 cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc
     4021 gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg
     4081 ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga
     4141 ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt
     4201 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     4261 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     4321 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     4381 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     4441 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     4501 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc
     4561 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat
     4621 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     4681 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
     4741 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     4801 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     4861 tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga
     4921 ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc
     4981 ggatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac
     5041 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca
     5101 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt
     5161 gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg
     5221 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga
     5281 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct
     5341 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg
     5401 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg
     5461 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc
     5521 gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat
     5581 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac
     5641 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt
     5701 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct
     5761 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc
     5821 tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca
     5881 ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga
     5941 tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag
     6001 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     6061 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac
     6121 cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
     6181 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     6241 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     6301 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     6361 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     6421 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     6481 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     6541 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     6601 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     6661 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     6721 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     6781 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     6841 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     6901 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     6961 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     7021 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     7081 ccgctggtag cggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     7141 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     7201 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     7261 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     7321 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     7381 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     7441 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     7501 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     7561 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     7621 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     7681 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     7741 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     7801 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     7861 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     7921 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     7981 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     8041 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     8101 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     8161 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     8221 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     8281 catttccccg aaaagtgcca cctgacgtc
//
