LOCUS       V042153                 7699 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V042153
VERSION     V042153
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"
     regulatory      913..922
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             928..1017
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     CDS             1033..3219
                     /label="Lrrfip1(NM_001111311)"
                     /note="Lrrfip1(NM_001111311)"
                     /gene="Lrrfip1"
     polyA_signal    3299..3523
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3569..3997
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4011..4340
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4407..5201
                     /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    5375..5496
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5545..5561)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5569..5585
                     /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(5593..5623)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5638..5659
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5947..6532)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6703..7563)
                     /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(7564..7668)
                     /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 aagccaccat ggcggcggaa caaaaactaa tatcagagga agatttggaa
      961 cagaaactaa tcagtgaaga agatttagag caaaagttaa tttccgagga ggacttatca
     1021 gcggaaaagc ttatgaccag cccagagggc gcgcaaaaca aggaaatcga ctgtctgagt
     1081 ccagaggcgc aaagactggc ggaggccagg ctcgcagcga agcgtgcagc ccgagccgag
     1141 gctcgagaga tccggatgaa ggagctggaa cggcagcaga aagaggtaga agaaagacca
     1201 gacaaagact ttgctgagaa gggctcccgt aacatgccta gcctgtctgc agccacactg
     1261 gcatctctgg gtgggacttc ctcccggaga gggagtggag atacctccat ctccatggac
     1321 accgaagcat ccatcaggga aatcaaggac tcgctggccg aagttgagga gaagtacaag
     1381 aaagctatgg tctccaacgc ccagctagac aatgagaaga ccaacttcat gtaccaggtg
     1441 gacacactga aggacatgct gctggagctg gaagagcagc tggccgagtc tcagcggcaa
     1501 tacgaggaga agaacaagga gttcgagagg gagaagcatg cccacagcat cctgcagttc
     1561 cagtttgccg aagtgaaaga ggccctgcgg caaagggaag agatgctcga gaaacatgga
     1621 ataatcctaa attcagaaat agccaccaac ggagagactt cagacacagt aaatgacgtt
     1681 gggtatcaag cccctacgaa gataacaaaa gaagagctga atgccctcaa gtcggccggg
     1741 gaggggaccc taggaaaagc caaagaagtg gaggtgaaaa aggaaattgt ggagaaagtg
     1801 gggcaaagag aaaccttaca gaattctgag caagaacagc ccaaacccaa cacaggaaag
     1861 gattgtgtgg acagaggggt gtcacatcct ggtgagaagg ctgagaacca gagacccgct
     1921 gaagacagtg ccctgtcccc gggaccacta gcaggggcca aatgtgagca acaggttcag
     1981 agccaagatc aagagaatac ttctgacctc aaaaactcag agcagatcga gtcacacaag
     2041 gtcacaaaca agtcagatag tagggccagt aactcccctg aacagtccag ctgcctggaa
     2101 ggtctagaca gtgaagtgcc agggcccact gaggatttga aaacagattt ggggaaaggt
     2161 agctttgaac catgtccaga ctacattttg gggcaaacag ctgaaattga caaggtgacc
     2221 tgtacagact caagaggcac aggtggaaac caaagggagg atgaggtaca ggcaggggac
     2281 accacagttg aggaccaagt agggacagtg gcttctggac ctgcaaagca aagcaaaggc
     2341 accgagaatc atggtgaaag ttgtctgaag gatgggctgg gacagagctc agagagagaa
     2401 ctcacccaag aagtagctga gcctgaggaa gccatcgttc agatcccaca ggccggtgga
     2461 gagaacacta ttaccaaggc agatgatgcc gaaggaaggg atgagaagcc catccaggca
     2521 gaagcccagg caagccctgg agctcccata aatcagagcg gccatcagga cacaacagga
     2581 ccaggcagca ccgatgccca gcgcacacca cctcatgcaa aagagcgcaa gaaacaaggg
     2641 aagagcgagc agcaggcaga ggccttggat tcaccgcaga agaagacgaa gaacaagaag
     2701 aagaagaaca agaagaagaa agctgcaact cccgcggaaa cctgcagaga tgccaatgaa
     2761 gagttgaact gtcaggaccc agatgtgggt gacatggagg aggaagagcg gctccaggtc
     2821 actgacaaga aacaggcatc aggaagccca gagcagaaaa tcagagcggg aagcagagaa
     2881 cctgtcgaag atcctcaaag tgggtcgagt ggaaaacaga acaaagtgga agaggatggt
     2941 cccacggaag gtcccacaga catcttggat cagaacagtc cccagtgtga agacagggag
     3001 atctccccag tgggagagaa aggtccccaa tgcgatacct ctcagatagg tagtgaagag
     3061 ggacatgtga cctcccaaca cggtggtcag gcagttgaga accacaacct agataacagt
     3121 gacctgtcag ggcagctgga gggcttcaat tcagaaagtg gagggcaggc aagagaggag
     3181 gttgggaata gcaagagtaa ggaggactgc accatgtcgt gagaattctg cagatatcca
     3241 gcacagtggc ggccgctcga gtctagaggg cccgtttaaa cccgctgatc agcctcgact
     3301 gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg
     3361 gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg
     3421 agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg
     3481 gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga
     3541 accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg
     3601 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     3661 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     3721 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     3781 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     3841 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     3901 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     3961 aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt
     4021 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     4081 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     4141 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     4201 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     4261 cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg
     4321 gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc
     4381 aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc
     4441 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct
     4501 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg
     4561 acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca
     4621 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc
     4681 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga
     4741 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc
     4801 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc
     4861 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg
     4921 ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct
     4981 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc
     5041 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc
     5101 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc
     5161 agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga
     5221 aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt
     5281 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
     5341 cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg
     5401 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     5461 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc
     5521 tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     5581 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     5641 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     5701 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     5761 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     5821 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     5881 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     5941 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     6001 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     6061 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     6121 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     6181 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     6241 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     6301 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     6361 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     6421 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     6481 cggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     6541 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     6601 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     6661 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     6721 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     6781 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     6841 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     6901 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     6961 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     7021 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     7081 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     7141 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     7201 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     7261 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     7321 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     7381 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     7441 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     7501 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     7561 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     7621 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     7681 aaaagtgcca cctgacgtc
//
