LOCUS       V035006                 6913 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V035006
VERSION     V035006
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..954
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             958..984
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             988..1014
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             1030..2433
                     /label="MIDN(NM_177401)"
                     /note="MIDN(NM_177401)"
                     /gene="MIDN"
     polyA_signal    2513..2737
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2783..3211
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3225..3554
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3621..4415
                     /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    4589..4710
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4759..4775)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4783..4799
                     /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(4807..4837)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4852..4873
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5161..5746)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5917..6777)
                     /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(6778..6882)
                     /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 ggcggcgtac ccatacgacg tacctgatta tgcaacttat
      961 ccttatgatg tacctgatta tgctagctat ccttatgatg tcccagacta cgcatcagcg
     1021 gaaaagctta tggagccgca gcccggcggc gcccggagct gccggcgcgg ggcccccggc
     1081 ggcgcctgcg agctgggccc ggcggccgag gcggcgccca tgagcctcgc catccacagc
     1141 accacgggca cccgctacga cctggccgtg ccgcccgacg agacggtgga ggggctgcgc
     1201 aagcggttgt cccagcgcct caaagtgccc aaggagcgcc tggctcttct ccacaaagac
     1261 acccggctca gttcggggaa gctgcaggag ttcggcgtgg gtgatggcag caagctgacc
     1321 ttggtaccca ccgtggaagc gggcctcatg tctcaggcct caaggccgga acagtccgtg
     1381 atgcaagctc tcgagagtct cacggagacg caggtcagtg acttcctgtc gggccgttcg
     1441 ccactgacac tggccttgcg tgtgggcgac cacatgatgt tcgtgcagct gcagctcgcg
     1501 gcccagcacg ctccactgca acaccgccat gtgctggccg ctgcggccgc cgccgctgct
     1561 gcgcgggggg accccagcat agcctccccc gtgtcctcgc cctgccggcc ggtgtccagt
     1621 gccgcccgag tccccccggt gcccaccagc ccgtcccctg catctccctc gcccatcaca
     1681 gccggctcct tccggtccca cgcagcctcc accacctgcc cggagcagat ggactgctcc
     1741 cccacggcca gcagcagtgc cagtcctggt gccagcacca cgtctacccc aggggccagc
     1801 cctgcccccc gctcccgaaa acccggcgcc gtcatcgaga gctttgtgaa tcacgccccg
     1861 ggggtcttct cagggacctt ctctggcacg ctacacccca actgccaaga cagcagcggg
     1921 cggccgcggc gtgacatcgg caccatcctg cagatcctga acgacctcct gagcgccacc
     1981 cggcactacc agggcatgcc cccttcgctg gcccagctcc gctgccacgc ccagtgctcc
     2041 ccggcctcac cggcccccga cctggccccc agaactacct cctgcgagaa gctcacggct
     2101 gccccctcag cctccctgct gcagggccag agccagatcc gcatgtgcaa gcccccgggg
     2161 gaccggcttc ggcagacaga aaaccgcgcc acgcgctgca aggtggaacg gctgcagctg
     2221 cttctgcagc agaaacggct ccgtagaaag gcccggcggg acgcgcgggg tccgtaccac
     2281 tggtcaccca gccgcaaggc cggccgcagc gacagcagta gcagcggggg cggcggcagc
     2341 cccagcgagg cctccggctt gggcctcgac ttcgaggact ccgtgtggaa gccagaagtc
     2401 aaccctgaca tcaagtcaga gttcgtggtg gcttaggaat tctgcagata tccagcacag
     2461 tggcggccgc tcgagtctag agggcccgtt taaacccgct gatcagcctc gactgtgcct
     2521 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt
     2581 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg
     2641 tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac
     2701 aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc
     2761 tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     2821 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     2881 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     2941 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     3001 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     3061 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     3121 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     3181 gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt
     3241 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     3301 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     3361 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     3421 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     3481 ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     3541 taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct gatcaagaga
     3601 caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg
     3661 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg
     3721 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt
     3781 ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg
     3841 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat
     3901 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat
     3961 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg
     4021 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg
     4081 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc
     4141 tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc
     4201 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
     4261 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg
     4321 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
     4381 tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac
     4441 cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga
     4501 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
     4561 tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa
     4621 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     4681 tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta
     4741 gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     4801 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     4861 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     4921 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     4981 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     5041 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     5101 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     5161 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     5221 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     5281 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     5341 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     5401 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     5461 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     5521 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     5581 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     5641 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggttt
     5701 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     5761 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     5821 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     5881 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     5941 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     6001 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     6061 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     6121 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     6181 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     6241 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     6301 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     6361 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     6421 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     6481 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     6541 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     6601 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     6661 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     6721 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     6781 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     6841 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     6901 gccacctgac gtc
//
