LOCUS       V022375                 7580 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V022375
VERSION     V022375
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..3037
                     /label="UVRAG(NM_003369)"
                     /note="UVRAG(NM_003369)"
                     /gene="UVRAG"
     CDS             3077..3142
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    3180..3404
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3450..3878
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3892..4221
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4288..5082
                     /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    5256..5377
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5426..5442)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5450..5466
                     /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(5474..5504)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5519..5540
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5828..6413)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6584..7444)
                     /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(7445..7549)
                     /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 atgagcgcct ccgcgtcggt
      961 cgggggcccc gtcccccagc cacccccggg cccggccgct gctctgcctc ccggttctgc
     1021 cgcgcgggcc ctgcatgtgg agctgccgtc tcagcagcgg cgtcttcgac atcttcggaa
     1081 cattgctgcc cggaacattg ttaatagaaa tggccatcag ctccttgata cctactttac
     1141 acttcacttg tgtagtactg aaaagatata taaagaattt tatagaagtg aagtgattaa
     1201 gaattccttg aatcccacgt ggcgaagtct cgattttgga attatgccag accgtcttga
     1261 tacatctgtg tcttgtttcg tggtgaagat atggggtgga aaggagaaca tctaccagct
     1321 gttgattgaa tggaaagtct gtttggatgg gctgaaatac ttgggtcagc agattcatgc
     1381 ccgaaaccaa aatgaaataa tttttgggct gaatgatgga tactatggtg ctccatttga
     1441 acataagggt tattcaaatg ctcagaagac tattcttctg caggtggatc agaactgtgt
     1501 tcgcaattct tacgatgtct tctctttgct acggcttcat agagcccagt gtgcaattaa
     1561 acagactcag gtaactgttc agaaaattgg aaaggaaatt gaagaaaaac taagactcac
     1621 atctacaagc aatgaactga aaaaaaaaag tgaatgcctg cagttaaaaa ttttggtgct
     1681 tcagaatgaa ctggaacggc agaagaaagc tttgggacgg gaggtggcat tactgcataa
     1741 gcaacaaatt gcattacaag acaaaggaag tgcattttca gctgagcacc tcaaacttca
     1801 actccagaag gaatccctaa atgagctgag gaaggagtgc actgcaaaaa gagaactctt
     1861 cttgaagact aatgctcagt tgacaattcg ttgcaggcag ttactctctg agctttccta
     1921 catttaccct attgatttga atgaacataa ggattacttt gtatgcggtg tcaagttgcc
     1981 taattctgag gacttccaag caaaagatga tggaagcatt gctgttgccc ttggttatac
     2041 tgcacatctg gtctccatga tttccttttt cctacaagtg cccctcagat atcctataat
     2101 tcataagggg tctagatcaa caatcaaaga caatatcaat gacaaactga cggaaaagga
     2161 gagagagttt ccactgtatc caaaaggagg ggagaagttg cagtttgatt atggtgtcta
     2221 tcttctgaac aaaaatatag cacagctaag atatcaacat ggactaggga ctccagactt
     2281 gcggcaaacc cttcccaacc tgaaaaactt catggagcat ggactaatgg tcaggtgtga
     2341 cagacatcac acctccagtg caatccctgt tcctaagaga caaagctcca tatttggggg
     2401 tgcagatgta ggcttctctg gggggatccc ttcaccagac aaaggacatc gaaaacgggc
     2461 cagctctgag aatgagagac ttcagtacaa aacccctcct cccagttaca actcagcatt
     2521 agcccagcct gtgaccaccg tcccctccat gggagagacc gagagaaaga taacatctct
     2581 atcctcctcc ttggatacct ccttggactt ctccaaagaa aacaagaaaa aaggagagga
     2641 tctagttggc agcttaaacg gaggccacgc gaatgtgcac cctagccaag aacaaggaga
     2701 agccctctcc gggcaccggg ccacagtcaa tggcactctc ctacccagcg agcaggccgg
     2761 gtccgccagt gtccagcttc caggcgagtt ccacccagtc tcagaagctg agctctgctg
     2821 tactgtggag caagcagaag aaatcatcgg gctggaagcc acaggtttcg cctcaggtga
     2881 tcagctagaa gcatttaact gcatcccagt ggacagtgct gtggcagtag agtgtgacga
     2941 acaagttctg ggagaatttg aagagttctc ccgaaggatc tatgcactga atgaaaacgt
     3001 atccagcttc cgccggccgc gcaggagttc cgataaggaa ttctgcagat atccagcaca
     3061 gtggcggccg ctcgaggact acaaagacca tgacggtgat tataaagatc atgacatcga
     3121 ctacaaggat gacgatgaca agtagtgagg gcccgtttaa acccgctgat cagcctcgac
     3181 tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct
     3241 ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct
     3301 gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg
     3361 ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag
     3421 aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc
     3481 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     3541 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     3601 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     3661 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     3721 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     3781 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
     3841 aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag
     3901 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     3961 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     4021 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     4081 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     4141 gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt
     4201 ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat
     4261 caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct
     4321 ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc
     4381 tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc
     4441 gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc
     4501 acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg
     4561 ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag
     4621 aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc
     4681 ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt
     4741 cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc
     4801 gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc
     4861 tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg
     4921 ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag
     4981 cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg
     5041 cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg
     5101 aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct
     5161 tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc
     5221 gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg
     5281 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     5341 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct
     5401 ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     5461 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     5521 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     5581 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     5641 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     5701 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     5761 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     5821 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     5881 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     5941 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     6001 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     6061 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     6121 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     6181 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     6241 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     6301 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     6361 gcggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     6421 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     6481 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     6541 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     6601 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     6661 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     6721 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     6781 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     6841 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     6901 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     6961 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     7021 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     7081 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     7141 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     7201 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     7261 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     7321 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     7381 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     7441 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     7501 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     7561 gaaaagtgcc acctgacgtc
//
