LOCUS       V033285                 7189 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V033285
VERSION     V033285
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             964..2679
                     /label="DPYSL4(NM_006426)"
                     /note="DPYSL4(NM_006426)"
                     /gene="DPYSL4"
     CDS             2686..2751
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    2789..3013
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3059..3487
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3501..3830
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3897..4691
                     /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    4865..4986
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5035..5051)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5059..5075
                     /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(5083..5113)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5128..5149
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5437..6022)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6193..7053)
                     /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(7054..7158)
                     /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 atccactagt ccagtgtggt ggaattcgcc
      961 accatgtcct tccagggcaa gaaaagcatc ccccggatca cgagtgaccg ccttctgatc
     1021 agaggtggga ggatcgtgaa tgacgaccag tccttttacg ctgatgtgca cgtggaagat
     1081 ggcttgataa aacaaatcgg agaaaacctc atcgtccctg ggggcatcaa gaccattgac
     1141 gcccacggcc tgatggtcct tcctggtggc gttgacgtcc acacaaggct gcagatgcct
     1201 gtcctgggca tgacaccggc tgacgacttc tgtcagggca ccaaggcagc gctagcagga
     1261 ggaaccacca tgatcttgga ccacgtcttc cccgacacgg gtgtgagcct gctggcggcc
     1321 tacgagcagt ggcgggagcg ggcggacagc gcggcctgct gcgactactc cctgcacgtg
     1381 gacatcaccc gatggcatga gagcatcaag gaggagctgg aggccctggt caaggagaag
     1441 ggtgtgaact ccttcctggt cttcatggca tacaaggacc ggtgccagtg cagcgacagc
     1501 cagatgtacg agatcttcag catcatccgg gacctggggg ccttggccca ggtgcacgct
     1561 gagaacgggg acatcgtgga ggaggagcag aagcggttgc tggagctcgg catcactggc
     1621 cccgagggcc acgtgctcag ccaccccgag gaggtggagg ctgaggcggt gtaccgagct
     1681 gtcaccatcg ccaagcaggc aaactgcccg ctgtacgtca ccaaggtgat gagcaagggg
     1741 gcggccgacg ccatcgctca ggccaagcgc agaggggtgg tcgtgtttgg ggagcccatc
     1801 accgccagcc tgggcaccga cggttcacac tactggagca agaactgggc caaggccgca
     1861 gccttcgtca catcaccccc tgtcaaccca gaccccacca cggcggacca cctcacctgc
     1921 ttgctgtcca gcggggacct ccaggtgaca ggcagcgccc actgcacctt caccactgcc
     1981 cagaaggctg tgggcaagga caacttcgcg ctgatccccg agggcaccaa cggcattgag
     2041 gagcgcatgt cgatggtctg ggagaaatgt gtggcctctg ggaagatgga cgagaatgag
     2101 ttcgtcgcgg tgaccagtac aaatgctgcc aaaatcttca atttttaccc aaggaagggg
     2161 cgagtggctg tgggctctga cgctgacctg gtcatatgga accccaaggc caccaagatc
     2221 atctctgcca agacccacaa tctgaacgtg gagtacaaca tcttcgaggg agtggagtgc
     2281 cggggagcgc ctgccgtggt cataagtcag ggccgagtgg cgctggagga cgggaagatg
     2341 tttgtcaccc cgggggcggg ccgcttcgtc cctcggaaaa cattcccgga ctttgtctac
     2401 aagaggatca aagctcgcaa caggctggcg gagatccacg gtgtgccccg tggactgtat
     2461 gacgggcccg tccacgaggt gatggtgcct gccaagccag ggagtggcgc tccggcccgc
     2521 gcgtcctgcc caggcaagat ctccgtccct cctgtgcgca acctacatca gtcggggttc
     2581 agcctatctg ggtctcaggc tgatgaccac atcgcccgac gcacagcaca gaagatcatg
     2641 gcaccacctg gcggccgctc caacatcacc tctctctccc tcgaggacta caaagaccat
     2701 gacggtgatt ataaagatca tgacatcgac tacaaggatg acgatgacaa gtagtgaggg
     2761 cccgtttaaa cccgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt
     2821 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa
     2881 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg
     2941 gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg
     3001 gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac
     3061 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     3121 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     3181 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     3241 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     3301 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     3361 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     3421 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     3481 gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     3541 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     3601 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     3661 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     3721 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc
     3781 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg
     3841 gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg tttcgcatga
     3901 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
     3961 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
     4021 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg
     4081 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
     4141 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
     4201 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
     4261 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
     4321 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
     4381 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg
     4441 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
     4501 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
     4561 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
     4621 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
     4681 agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc
     4741 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     4801 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     4861 ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     4921 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     4981 atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata
     5041 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     5101 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     5161 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     5221 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     5281 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     5341 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     5401 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     5461 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     5521 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     5581 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     5641 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     5701 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     5761 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     5821 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     5881 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     5941 ttgatccggc aaacaaacca ccgctggtag cggttttttt gtttgcaagc agcagattac
     6001 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     6061 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     6121 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     6181 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     6241 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     6301 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     6361 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     6421 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     6481 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     6541 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     6601 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     6661 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     6721 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     6781 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     6841 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     6901 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     6961 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     7021 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     7081 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     7141 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtc
//
