LOCUS       V030195                 7619 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V030195
VERSION     V030195
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..3109
                     /label="LRSAM1(NM_138361)"
                     /note="LRSAM1(NM_138361)"
                     /gene="LRSAM1"
     CDS             3116..3181
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    3219..3443
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3489..3917
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3931..4260
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4327..5121
                     /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    5295..5416
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5465..5481)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5489..5505
                     /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(5513..5543)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5558..5579
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5867..6452)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6623..7483)
                     /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(7484..7588)
                     /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 atgccgctct tcttccggaa
      961 gcggaaaccc agtgaggagg ctcggaaacg cctggagtac cagatgtgtt tggcaaaaga
     1021 agctggggca gatgacattc tcgacatctc taaatgtgag ctctcagaga ttccatttgg
     1081 agcttttgca acatgcaaag ttctgcagaa gaaggtgctg atcgtccaca cgaatcacct
     1141 cacttccctg cttcccaaat cctgcagcct cctgagtctg gcaaccatca aggttctaga
     1201 tctccacgat aatcagctga cagcccttcc tgacgatctg gggcagctga ctgccctcca
     1261 ggtcttaaac gtggaaagga atcaactgat gcagctccca cgttccattg ggaacctgac
     1321 ccagctccag actctcaatg ttaaagacaa caagctgaag gagcttccag acaccgtggg
     1381 ggagcttcga agcctgcgta ccctcaacat cagtggaaac gagatccaga gattgccgca
     1441 gatgctggct cacgttcgaa ccctggagat gctgagcctt gacgcctcgg ccatggtcta
     1501 cccgccgcgg gaggtgtgtg gtgccggcac tgcggccatc ttgcagttcc tctgcaaaga
     1561 gtcagggctg gaatactacc ccccttctca gtacttgctg ccaattctgg agcaagatgg
     1621 aatcgagaac tctcgggaca gccctgatgg gcccacggac agattctcaa gggaggagtt
     1681 agagtggcag aacaggttct cagactatga gaagaggaag gaacagaaga tgctggagaa
     1741 actcgagttt gaacggcgcc tggaactggg gcagcgggag cacacccagc tccttcagca
     1801 gagcagcagc cagaaggatg agatccttca gacggtcaag gaggagcagt cccggctgga
     1861 gcagggcctg agtgagcacc agcgccacct caacgcagag cggcagcggc tgcaggagca
     1921 gctgaagcag acggaacaga acatttccag ccggatccag aagctgctgc aggacaatca
     1981 gagacaaaag aaaagctccg agattttgaa atcgctggaa aatgaaagaa taagaatgga
     2041 acagttgatg tccataaccc aggaggagac tgagagcctg cggcgacgtg acgttgcctc
     2101 cgccatgcag cagatgctga ctgagagctg taagaaccgg ctcatccaga tggcctacga
     2161 atctcagagg cagaacttgg tccagcaggc ctgttccagc atggccgaaa tggatgaacg
     2221 attccagcag attctgtcgt ggcagcaaat ggatcagaac aaagccatca gccagatcct
     2281 gcaggagagc gcgatgcaga aggctgcgtt cgaggcactc caggtgaaga aagacctgat
     2341 gcatcggcag atcaggagcc agattaagtt aatagaaact gagttattgc agctgacaca
     2401 gctggagtta aagaggaagt ccctggacac agagtcactc caggagatga tctcggagca
     2461 gcgctgggcc ctcagctccc tgctccagca gctgctcaaa gagaagcagc agcgagagga
     2521 agagctccgg gaaatcctga cggagttaga agccaaaagt gaaaccaggc aggaaaatta
     2581 ctggctgatt cagtatcaac ggcttttgaa ccagaagccc ttgtccttga agctgcaaga
     2641 agaggggatg gagcgccagc tggtggccct cctggaggag ctgtcggctg agcactacct
     2701 gcccatcttt gcgcaccacc gcctctcact ggacctgctg agccaaatga gcccagggga
     2761 cctggccaag gtgggcgtct cagaagctgg cctgcagcac gagatcctcc ggagagtcca
     2821 ggaactgctg gatgcagcca ggatccagcc agagctgaaa ccaccaatgg gtgaggtcgt
     2881 cacccctacg gccccccagg agcctcctga gtctgtgagg ccatccgctc cccctgcaga
     2941 gctggaggtg caggcctcag agtgtgtcgt gtgcctggaa cgggaggccc agatgatctt
     3001 cctcaactgt ggccacgtct gctgctgcca gcagtgctgc cagccactgc gcacctgccc
     3061 gctgtgccgc caggacatcg cccagcgcct ccgcatctac cacagcagcc tcgaggacta
     3121 caaagaccat gacggtgatt ataaagatca tgacatcgac tacaaggatg acgatgacaa
     3181 gtagtgaggg cccgtttaaa cccgctgatc agcctcgact gtgccttcta gttgccagcc
     3241 atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt
     3301 cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct
     3361 ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc
     3421 tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg
     3481 gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     3541 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     3601 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     3661 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     3721 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     3781 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     3841 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
     3901 aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
     3961 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
     4021 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     4081 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     4141 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct
     4201 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     4261 aagctcccgg gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg
     4321 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     4381 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     4441 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     4501 gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     4561 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     4621 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     4681 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     4741 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     4801 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg
     4861 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     4921 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     4981 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     5041 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     5101 cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc
     5161 ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg
     5221 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt
     5281 tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     5341 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     5401 tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat
     5461 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     5521 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     5581 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     5641 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     5701 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     5761 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     5821 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     5881 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     5941 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     6001 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     6061 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     6121 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     6181 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     6241 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     6301 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     6361 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggttttttt gtttgcaagc
     6421 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     6481 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     6541 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     6601 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     6661 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     6721 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     6781 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     6841 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     6901 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     6961 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     7021 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     7081 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     7141 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     7201 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     7261 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     7321 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     7381 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     7441 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     7501 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     7561 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtc
//
