LOCUS       V026189                 7099 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V026189
VERSION     V026189
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             922..987
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     CDS             991..2619
                     /label="EHD2(NM_014601)"
                     /note="EHD2(NM_014601)"
                     /gene="EHD2"
     polyA_signal    2699..2923
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2969..3397
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3411..3740
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3807..4601
                     /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    4775..4896
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4945..4961)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4969..4985
                     /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(4993..5023)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5038..5059
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5347..5932)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6103..6963)
                     /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(6964..7068)
                     /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 ggactacaaa gaccatgacg gtgattataa agatcatgac
      961 atcgactaca aggatgacga tgacaagctt atgttcagct ggctgaagcg gggcggggca
     1021 cggggccagc agcccgaggc catccgcacg gtgacctcgg ccctcaagga gctgtaccgc
     1081 acgaagctgc tgccgctgga ggagcactac cgctttgggg ccttccactc gccggccctg
     1141 gaggacgcag acttcgacgg caagcccatg gtgctggtgg ccggccagta cagcacgggc
     1201 aagaccagct tcatccagta cctgctggag caggaggtgc ccggctcccg cgtggggcct
     1261 gagcccacca ccgactgctt tgtggccgtc atgcacgggg acactgaggg caccgtgccc
     1321 ggcaacgccc tcgtcgtgga cccggacaag cccttccgca aactcaaccc tttcggaaac
     1381 accttcctca acaggttcat gtgtgcccag ctccctaatc aggtcctgga gagcatcagc
     1441 atcatcgaca ccccgggtat cctgtcgggt gccaagcaga gagtgagccg cggctacgac
     1501 ttcccggccg tgctgcgctg gttcgcggag cgcgtggacc tcatcatcct gctctttgat
     1561 gcgcacaagc tggagatctc ggacgagttc tcagaggcca tcggcgcgtt gcggggccat
     1621 gaggacaaga tccgcgtggt gctcaacaag gccgacatgg tggagacgca gcagctgatg
     1681 cgcgtctacg gcgcgctcat gtgggcgctg ggcaaggtgg tgggcacgcc cgaggtgctg
     1741 cgcgtctaca tcggctcctt ctggtcccag cccctcctcg tgcccgacaa ccggcgcctc
     1801 ttcgagctgg aggagcagga cctcttccgc gacatccagg gcctgccccg gcacgcagcc
     1861 ttgcgcaagc tcaacgacct ggtgaagagg gcccggctgg tgcgagttca cgcttacatc
     1921 atcagctacc tgaagaagga gatgccctct gtgtttggga aggagaacaa gaagaagcag
     1981 ctgatcctca aactgcccgt catctttgcg aagattcagc tggaacatca catctcccct
     2041 ggggactttc ctgattgcca gaaaatgcag gagctgctga tggcgcacga cttcaccaag
     2101 tttcactcgc tgaagccgaa gctgctagag gcactggacg agatgctgac gcacgacatc
     2161 gccaagctca tgcccctgct gcggcaggag gagctggaga gcaccgaggt gggcgtgcag
     2221 gggggcgctt ttgagggcac ccacatgggc ccgtttgtgg agcggggacc tgacgaggcc
     2281 atggaggacg gcgaggaggg ctcggacgac gaggccgagt gggtggtgac caaggacaag
     2341 tccaaatacg acgagatctt ctacaacctg gcgcctgccg acggcaagct gagcggctcc
     2401 aaggccaaga cctggatggt ggggaccaag ctccccaact cagtgctggg gcgcatctgg
     2461 aagctcagcg atgtggaccg cgacggcatg ctggatgatg aggagttcgc gctggccagc
     2521 cacctcatcg aggccaagct ggaaggccac gggctgcccg ccaacctgcc ccgtcgcctg
     2581 gtgccaccct ccaagcgacg ccacaagggc tccgccgagt gagaattctg cagatatcca
     2641 gcacagtggc ggccgctcga gtctagaggg cccgtttaaa cccgctgatc agcctcgact
     2701 gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg
     2761 gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg
     2821 agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg
     2881 gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga
     2941 accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg
     3001 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     3061 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     3121 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     3181 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     3241 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     3301 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     3361 aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt
     3421 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     3481 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     3541 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     3601 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     3661 cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg
     3721 gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc
     3781 aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc
     3841 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct
     3901 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg
     3961 acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca
     4021 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc
     4081 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga
     4141 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc
     4201 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc
     4261 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg
     4321 ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct
     4381 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc
     4441 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc
     4501 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc
     4561 agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga
     4621 aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt
     4681 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
     4741 cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg
     4801 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     4861 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc
     4921 tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     4981 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     5041 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     5101 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     5161 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     5221 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     5281 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     5341 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     5401 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     5461 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     5521 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     5581 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     5641 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     5701 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     5761 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     5821 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     5881 cggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     5941 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     6001 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     6061 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     6121 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     6181 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     6241 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     6301 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     6361 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     6421 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     6481 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     6541 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     6601 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     6661 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     6721 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     6781 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     6841 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     6901 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     6961 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     7021 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     7081 aaaagtgcca cctgacgtc
//
