LOCUS       V026488                 7759 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V026488
VERSION     V026488
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..3279
                     /label="Tfrc(NM_011638)"
                     /note="Tfrc(NM_011638)"
                     /gene="Tfrc"
     polyA_signal    3359..3583
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3629..4057
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4071..4400
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4467..5261
                     /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    5435..5556
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5605..5621)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5629..5645
                     /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(5653..5683)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5698..5719
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6007..6592)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6763..7623)
                     /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(7624..7728)
                     /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 atgatggatc aagccagatc agcattctct
     1021 aacttgtttg gtggggaacc attgtcatac acccggttta gccttgctcg gcaagtagat
     1081 ggagataaca gtcatgtgga gatgaaactg gctgcagatg aagaagaaaa tgccgacaat
     1141 aacatgaagg ctagtgtcag aaaacccaag aggtttaatg gaagactctg ctttgcagct
     1201 attgcactag tcattttctt cttgattgga ttcatgagtg gctacctggg ctattgtaag
     1261 cgtgtagaac aaaaagagga gtgtgtgaaa ctggctgaaa cggaggagac agacaagtca
     1321 gaaaccatgg aaacagagga tgttcctaca tcatctcgct tatattgggc agacctcaaa
     1381 acactgttgt cagagaagtt gaactccata gagtttgctg acaccatcaa gcagctgagc
     1441 cagaatacat acactcctcg tgaggctgga tctcaaaaag atgaaagtct tgcctattat
     1501 attgaaaatc agttccatga atttaaattc agcaaagtct ggcgagatga acactatgtg
     1561 aagattcaag tgaaaagcag cattggtcaa aacatggtga ccatagtgca gtcaaatggt
     1621 aacttagacc cagtggagtc tcccgagggt tatgtggcat tcagtaaacc tacagaagtt
     1681 tctggtaaac tggtccatgc taattttggc actaaaaagg actttgaaga actaagttat
     1741 tctgtgaatg gatctttagt gattgttaga gcaggggaaa ttacttttgc agaaaaggtt
     1801 gcaaatgccc aaagctttaa tgcaattggt gtcctcatat acatggacaa gaataaattc
     1861 cccgttgttg aggcagacct tgcactcttt ggacatgctc atctaggaac tggtgatcca
     1921 tacacacctg gctttccttc tttcaatcat actcagtttc cgccatctca gtcatcaggg
     1981 ttgcctaata tacctgtgca aacaatctca agagctgctg cagaaaagct atttggaaaa
     2041 atggaaggaa gctgtcctgc tagatggaac atagattctt catgtaagct ggaactttca
     2101 cagaatcaaa atgtgaagct cattgtgaaa aacgtactga aagaaagaag aatacttaac
     2161 atctttggag ttattaaagg ttatgaggaa ccagaccgtt atgttgtagt aggagcccag
     2221 agagacgctt tgggtgctgg tgttgcggcg aagtccagtg tgggaacagg tcttctgttg
     2281 aaacttgccc aagtattctc agatatgatt tcaaaagatg gatttagacc cagcagaagt
     2341 ataatctttg ccagctggac tgcaggcgac tttggagctg ttggtgccac tgagtggttg
     2401 gagggatacc tttcatcttt gcatttaaaa gctttcactt atattaattt ggataaagtt
     2461 gtccttggta ctagtaactt caaagtttct gccagcccct tattatatac acttatggga
     2521 aagataatgc aagatgtaaa gcatccagtt gatggaaaat ctctatatag agacagcaat
     2581 tggattagca aagttgagaa actttccttt gacaatgctg catatccttt ccttgcatat
     2641 tctggaatcc cagcagtttc tttttgtttt tgtgaggatg cagactatcc ttatttgggc
     2701 actagattgg atacctatga ggcattgact cagaaagttc ctcagctcaa ccaaatggtt
     2761 cgtacagcag cggaagtggc tggtcagctc attattaaac ttacccatga cgttgaattg
     2821 aacctggact atgagatgta taacagcaaa ctactgtcat ttatgaagga tctgaaccag
     2881 ttcaaaacag atatcaggga tatgggtcta agtctacagt ggctgtattc cgctcgtgga
     2941 gactacttcc gtgctacttc tagactaaca actgattttc ataatgctga gaaaacaaac
     3001 agatttgtca tgagggaaat caatgatcgt attatgaaag tggagtatca cttcctgtcg
     3061 ccctatgtat ctccaagaga gtctcctttc cgacatatct tctggggctc tggctctcac
     3121 actctctcag ctttagtgga gaacttgaag cttcgtcaaa aaaatattac tgcttttaat
     3181 gaaaccctct tcagaaacca gttggccctg gctacttgga ctattcaggg agtcgcaaat
     3241 gccctctctg gtgacatttg gaatattgac aatgagtttt aagaattctg cagatatcca
     3301 gcacagtggc ggccgctcga gtctagaggg cccgtttaaa cccgctgatc agcctcgact
     3361 gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg
     3421 gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg
     3481 agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg
     3541 gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga
     3601 accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg
     3661 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     3721 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     3781 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     3841 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     3901 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     3961 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     4021 aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt
     4081 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     4141 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     4201 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     4261 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     4321 cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg
     4381 gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc
     4441 aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc
     4501 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct
     4561 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg
     4621 acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca
     4681 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc
     4741 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga
     4801 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc
     4861 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc
     4921 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg
     4981 ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct
     5041 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc
     5101 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc
     5161 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc
     5221 agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga
     5281 aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt
     5341 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
     5401 cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg
     5461 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     5521 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc
     5581 tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     5641 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     5701 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     5761 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     5821 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     5881 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     5941 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     6001 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     6061 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     6121 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     6181 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     6241 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     6301 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     6361 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     6421 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     6481 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     6541 cggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     6601 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     6661 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     6721 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     6781 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     6841 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     6901 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     6961 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     7021 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     7081 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     7141 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     7201 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     7261 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     7321 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     7381 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     7441 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     7501 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     7561 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     7621 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     7681 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     7741 aaaagtgcca cctgacgtc
//
