LOCUS       V035285                 7488 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V035285
VERSION     V035285
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        50..429
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        430..633
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        678..696
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             756..2168
                     /label="Tymp(NM_138302)"
                     /note="Tymp(NM_138302)"
                     /gene="Tymp"
     CDS             2175..2240
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2264..2852
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2918..2966
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3168..3596
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3610..3939
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4006..4800
                     /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    4976..5097
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5146..5162)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5170..5186
                     /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(5194..5224)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5239..5260
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5548..6136)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6307..7167)
                     /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(7168..7272)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta
       61 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
      121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      421 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      601 cgtgtacggt gggaggtcta tataagcaga gctctctggc taactagaga acccactgct
      661 tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagcgttta
      721 aacttaagct tggtaccgag ctcgaattcg ccaccatggc agctccaggg acaccacctc
      781 cctcggcctc agggggagga ggtggggagc ccaggcagct gcctgagctg atccgcctga
      841 agcgagatgg agggcatctg agagaagcag atatcagaaa cttcgtgcac gcggtgatag
      901 atggaagagc acaggacaca caaattgggg ccatgctaat ggccattcgg ctgcagggaa
      961 tgaacctaga agagacatct gtgctgactc gggcccttgc agagtctggg cagcaactgg
     1021 agtggcccaa agcctggcac cagcagcttg ttgacaaaca ctccacagga ggtgtgggtg
     1081 acaaggttag cctggttctg gcacctgccc tggctgcatg cggctgtaag gtgccaatga
     1141 tcagcggccg cagtctggga cacacagggg gcacactgga taagctggaa tcgattcctg
     1201 ggttcggtgt cacccagagc ccagagcaga tgctgcacat ccttgaggaa gtcggctgct
     1261 gcattgtcgg ccagagtgca aagctggttc ctgctgatgg gatcctgtat gctgcacgag
     1321 atgtgacagc cactgtggac agtgtcccac tcattacagc ttcaatcctc agtaagaagg
     1381 ccgtggaggg actgtccact ctggtggtag atgttaagtt tgggggagct gcagtcttcc
     1441 cagaccagga gaaggcccga gaactggcaa agatgttggt tagagtggga gtgagcctgg
     1501 ggctgaaggt cgccgcagcc ctgaccgcca tggataaccc tctgggccgc agcgtgggcc
     1561 ataccctgga ggtggaagaa gcgttgcttt gcctggatgg tgcggggccg ccggatctgc
     1621 gggacctggt cattaggcta ggaggcgcca ttctttggat tagcggacag gcggaaacac
     1681 aagaccaagg cgccgcccga gtggccgcgg cgttggatga cggctccgcg cggcgccgct
     1741 tccagctgat gctttcggcg cagggcgtgg accccggcct agctaaagca ttgtgctccg
     1801 ggagccccac gcaacgccgg cagctgctgc ctcacgcccg agagcaagag gagctgctcg
     1861 caccggcaga cggcattgtt gagtgcgtcc gagcgctgcc gctggcccgt gtgctgcacg
     1921 acctcggagc cggacgcagc cgagcggggc agccgatcag accgggagtg ggtgccgagg
     1981 tactggtcga cgtgggccag tgtttgtccc gagggacacc ctggctccgc gtgcacctag
     2041 acggccctgc gctcagcagt cagcagcgac gcactctcca gggggcgctc gtgctatcag
     2101 atcgagcacc cttcaaggtc ccttcaccct tcgctgaact tgtcctccca ccgaccatcg
     2161 cacagcccgg atccgactac aaagaccatg acggtgatta taaagatcat gacatcgact
     2221 acaaggatga cgatgacaag tagtgagggc ccgatatctc gacaatcaac ctctggatta
     2281 caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg
     2341 atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc
     2401 ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca
     2461 acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac
     2521 cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact
     2581 catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc
     2641 cgtggtgttg tcggggaagc tgacgtcctt tccatggctg ctcgcctgtg ttgccacctg
     2701 gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc
     2761 ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac
     2821 gagtcggatc tccctttggg ccgcctcccc gcctggaaac gggggaggct aactgaaaca
     2881 cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa
     2941 acgcacgggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc
     3001 tgtcgatacc ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc
     3061 accccacccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg
     3121 ccctgccata gcagatctgc gcagctgggg ctctaggggg tatccccacg cgccctgtag
     3181 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     3241 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     3301 tccccgtcaa gctctaaatc ggggcatccc tttagggttc cgatttagtg ctttacggca
     3361 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     3421 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     3481 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttggg
     3541 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt
     3601 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     3661 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     3721 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     3781 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     3841 ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag
     3901 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata
     3961 tccattttcg gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat
     4021 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     4081 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     4141 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg
     4201 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     4261 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     4321 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     4381 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     4441 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     4501 gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc
     4561 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     4621 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     4681 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     4741 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     4801 gcgggactct ggggttcgcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga
     4861 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc
     4921 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt
     4981 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     5041 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
     5101 tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc
     5161 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     5221 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     5281 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     5341 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     5401 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     5461 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     5521 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     5581 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     5641 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     5701 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct caatgctcac gctgtaggta
     5761 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     5821 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     5881 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     5941 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg
     6001 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     6061 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     6121 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     6181 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     6241 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     6301 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     6361 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     6421 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     6481 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     6541 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     6601 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     6661 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     6721 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     6781 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     6841 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     6901 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     6961 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     7021 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     7081 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     7141 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     7201 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     7261 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtcgacggat cgggagatct
     7321 cccgatcccc tatggtcgac tctcagtaca atctgctctg atgccgcata gttaagccag
     7381 tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa aatttaagct
     7441 acaacaaggc aaggcttgac cgacaattgc atgaagaatc tgcttagg
//
