LOCUS       V046833                 6868 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V046833
VERSION     V046833
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..1527
                     /label="GCLM(NM_002061)"
                     /note="GCLM(NM_002061)"
                     /gene="GCLM"
     CDS             1534..1560
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             1564..1590
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             1594..1620
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     misc_feature    1644..2232
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2298..2346
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      2548..2976
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        2990..3319
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3386..4180
                     /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    4356..4477
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4526..4542)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4550..4566
                     /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(4574..4604)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4619..4640
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4928..5516)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5687..6547)
                     /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(6548..6652)
                     /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 ccaccatggg caccgacagc
      721 cgcgcggcca aggcgctcct ggcgcgggcc cgcaccctgc acctgcagac ggggaacctg
      781 ctgaactggg gccgcctgcg gaagaagtgc ccgtccacgc acagcgagga gcttcatgat
      841 tgtatccaaa aaaccttgaa tgaatggagt tcccaaatca acccagattt ggtcagggag
      901 tttccagatg tcttggaatg cactgtatct catgcagtag aaaagataaa tcctgatgaa
      961 agagaagaaa tgaaagtttc tgcaaaactg ttcattgtag aatcaaactc ttcatcatca
     1021 actagaagtg cagttgacat ggcctgttca gtccttggag ttgcacagct ggattctgtg
     1081 atcattgctt cacctcctat tgaagatgga gttaatcttt ccttggagca tttacagcct
     1141 tactgggagg aattagaaaa cttagttcag agcaaaaaga ttgttgccat aggtacctct
     1201 gatctagaca aaacacagtt ggaacagctg tatcagtggg cacaggtaaa accaaatagt
     1261 aaccaagtta atcttgcctc ctgctgtgtg atgccaccag atttgactgc atttgctaaa
     1321 caatttgaca tacagctgtt gactcacaat gatccaaaag aactgctttc tgaagcaagt
     1381 ttccaagaag ctcttcagga aagcattcct gacattcaag cgcacgagtg ggtgccgctg
     1441 tggctactgc ggtattcggt cattgtgaaa agtagaggaa ttatcaaatc aaaaggctac
     1501 attttacaag ctaaaagaag gggttctctc gagtacccat acgacgtacc tgattatgca
     1561 acttatcctt atgatgtacc tgattatgct agctatcctt atgatgtccc agactacgca
     1621 tagtgagggc ccgatatctc gacaatcaac ctctggatta caaaatttgt gaaagattga
     1681 ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt
     1741 tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt
     1801 tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg
     1861 tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg
     1921 ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc
     1981 gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaagc
     2041 tgacgtcctt tccatggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct
     2101 tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg
     2161 ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg
     2221 ccgcctcccc gcctggaaac gggggaggct aactgaaaca cggaaggaga caataccgga
     2281 aggaacccgc gctatgacgg caataaaaag acagaataaa acgcacgggt gttgggtcgt
     2341 ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac
     2401 cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg
     2461 gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata gcagatctgc
     2521 gcagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg
     2581 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
     2641 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     2701 ggggcatccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     2761 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     2821 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
     2881 ctatctcggt ctattctttt gatttataag ggattttggg gatttcggcc tattggttaa
     2941 aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt
     3001 agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     3061 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     3121 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct
     3181 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     3241 agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg
     3301 aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca
     3361 agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc
     3421 ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc
     3481 tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga
     3541 cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac
     3601 gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct
     3661 gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa
     3721 agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc
     3781 attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct
     3841 tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc
     3901 caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg
     3961 cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct
     4021 gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct
     4081 tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca
     4141 gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgcg
     4201 aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct
     4261 tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc
     4321 gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg
     4381 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     4441 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct
     4501 ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     4561 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     4621 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     4681 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     4741 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     4801 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     4861 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     4921 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     4981 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     5041 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     5101 cgggaagcgt ggcgctttct caatgctcac gctgtaggta tctcagttcg gtgtaggtcg
     5161 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     5221 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     5281 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     5341 ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc
     5401 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     5461 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     5521 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     5581 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     5641 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     5701 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     5761 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     5821 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     5881 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     5941 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     6001 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     6061 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     6121 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     6181 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     6241 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     6301 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     6361 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     6421 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     6481 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     6541 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     6601 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     6661 cccgaaaagt gccacctgac gtcgacggat cgggagatct cccgatcccc tatggtcgac
     6721 tctcagtaca atctgctctg atgccgcata gttaagccag tatctgctcc ctgcttgtgt
     6781 gttggaggtc gctgagtagt gcgcgagcaa aatttaagct acaacaaggc aaggcttgac
     6841 cgacaattgc atgaagaatc tgcttagg
//
