LOCUS       V031547                 8008 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V031547
VERSION     V031547
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"
     CDS             706..2667
                     /label="HSPA5(NM_005347)"
                     /note="HSPA5(NM_005347)"
                     /gene="HSPA5"
     CDS             2674..2700
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             2704..2730
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             2734..2760
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     misc_feature    2784..3372
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3438..3486
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3688..4116
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4130..4459
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4526..5320
                     /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    5496..5617
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5666..5682)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5690..5706
                     /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(5714..5744)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5759..5780
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6068..6656)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6827..7687)
                     /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(7688..7792)
                     /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 gctctggcta gcgtttaaac ttaagcttgg
      661 taccgagctc ggatccacta gtccagtgtg gtggaattcg ccaccatgaa gctctccctg
      721 gtggccgcga tgctgctgct gctcagcgcg gcgcgggccg aggaggagga caagaaggag
      781 gacgtgggca cggtggtcgg catcgacctg gggaccacct actcctgcgt cggcgtgttc
      841 aagaacggcc gcgtggagat catcgccaac gatcagggca accgcatcac gccgtcctat
      901 gtcgccttca ctcctgaagg ggaacgtctg attggcgatg ccgccaagaa ccagctcacc
      961 tccaaccccg agaacacggt ctttgacgcc aagcggctca tcggccgcac gtggaatgac
     1021 ccgtctgtgc agcaggacat caagttcttg ccgttcaagg tggttgaaaa gaaaactaaa
     1081 ccatacattc aagttgatat tggaggtggg caaacaaaga catttgctcc tgaagaaatt
     1141 tctgccatgg ttctcactaa aatgaaagaa accgctgagg cttatttggg aaagaaggtt
     1201 acccatgcag ttgttactgt accagcctat tttaatgatg cccaacgcca agcaaccaaa
     1261 gacgctggaa ctattgctgg cctaaatgtt atgaggatca tcaacgagcc tacggcagct
     1321 gctattgctt atggcctgga taagagggag ggggagaaga acatcctggt gtttgacctg
     1381 ggtggcggaa ccttcgatgt gtctcttctc accattgaca atggtgtctt cgaagttgtg
     1441 gccactaatg gagatactca tctgggtgga gaagactttg accagcgtgt catggaacac
     1501 ttcatcaaac tgtacaaaaa gaagacgggc aaagatgtca ggaaagacaa tagagctgtg
     1561 cagaaactcc ggcgcgaggt agaaaaggcc aaacgggccc tgtcttctca gcatcaagca
     1621 agaattgaaa ttgagtcctt ctatgaagga gaagactttt ctgagaccct gactcgggcc
     1681 aaatttgaag agctcaacat ggatctgttc cggtctacta tgaagcccgt ccagaaagtg
     1741 ttggaagatt ctgatttgaa gaagtctgat attgatgaaa ttgttcttgt tggtggctcg
     1801 actcgaattc caaagattca gcaactggtt aaagagttct tcaatggcaa ggaaccatcc
     1861 cgtggcataa acccagatga agctgtagcg tatggtgctg ctgtccaggc tggtgtgctc
     1921 tctggtgatc aagatacagg tgacctggta ctgcttgatg tatgtcccct tacacttggt
     1981 attgaaactg tgggaggtgt catgaccaaa ctgattccaa ggaacacagt ggtgcctacc
     2041 aagaagtctc agatcttttc tacagcttct gataatcaac caactgttac aatcaaggtc
     2101 tatgaaggtg aaagacccct gacaaaagac aatcatcttc tgggtacatt tgatctgact
     2161 ggaattcctc ctgctcctcg tggggtccca cagattgaag tcacctttga gatagatgtg
     2221 aatggtattc ttcgagtgac agctgaagac aagggtacag ggaacaaaaa taagatcaca
     2281 atcaccaatg accagaatcg cctgacacct gaagaaatcg aaaggatggt taatgatgct
     2341 gagaagtttg ctgaggaaga caaaaagctc aaggagcgca ttgatactag aaatgagttg
     2401 gaaagctatg cctattctct aaagaatcag attggagata aagaaaagct gggaggtaaa
     2461 ctttcctctg aagataagga gaccatggaa aaagctgtag aagaaaagat tgaatggctg
     2521 gaaagccacc aagatgctga cattgaagac ttcaaagcta agaagaagga actggaagaa
     2581 attgttcaac caattatcag caaactctat ggaagtgcag gccctccccc aactggtgaa
     2641 gaggatacag cagaaaaaga tgagttgctc gagtacccat acgacgtacc tgattatgca
     2701 acttatcctt atgatgtacc tgattatgct agctatcctt atgatgtccc agactacgca
     2761 tagtgagggc ccgatatctc gacaatcaac ctctggatta caaaatttgt gaaagattga
     2821 ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt
     2881 tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt
     2941 tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg
     3001 tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg
     3061 ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc
     3121 gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaagc
     3181 tgacgtcctt tccatggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct
     3241 tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg
     3301 ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg
     3361 ccgcctcccc gcctggaaac gggggaggct aactgaaaca cggaaggaga caataccgga
     3421 aggaacccgc gctatgacgg caataaaaag acagaataaa acgcacgggt gttgggtcgt
     3481 ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac
     3541 cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg
     3601 gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata gcagatctgc
     3661 gcagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg
     3721 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
     3781 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     3841 ggggcatccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     3901 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     3961 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
     4021 ctatctcggt ctattctttt gatttataag ggattttggg gatttcggcc tattggttaa
     4081 aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt
     4141 agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     4201 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     4261 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct
     4321 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     4381 agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg
     4441 aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca
     4501 agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc
     4561 ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc
     4621 tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga
     4681 cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac
     4741 gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct
     4801 gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa
     4861 agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc
     4921 attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct
     4981 tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc
     5041 caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg
     5101 cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct
     5161 gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct
     5221 tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca
     5281 gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgcg
     5341 aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct
     5401 tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc
     5461 gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg
     5521 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     5581 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct
     5641 ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     5701 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     5761 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     5821 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     5881 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     5941 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     6001 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     6061 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     6121 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     6181 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     6241 cgggaagcgt ggcgctttct caatgctcac gctgtaggta tctcagttcg gtgtaggtcg
     6301 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     6361 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     6421 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     6481 ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc
     6541 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     6601 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     6661 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     6721 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     6781 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     6841 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     6901 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     6961 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     7021 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     7081 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     7141 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     7201 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     7261 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     7321 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     7381 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     7441 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     7501 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     7561 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     7621 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     7681 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     7741 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     7801 cccgaaaagt gccacctgac gtcgacggat cgggagatct cccgatcccc tatggtcgac
     7861 tctcagtaca atctgctctg atgccgcata gttaagccag tatctgctcc ctgcttgtgt
     7921 gttggaggtc gctgagtagt gcgcgagcaa aatttaagct acaacaaggc aaggcttgac
     7981 cgacaattgc atgaagaatc tgcttagg
//
