LOCUS       V045216                10338 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V045216
VERSION     V045216
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..5012
                     /label="FMN1(NM_001277313)"
                     /note="FMN1(NM_001277313)"
                     /gene="FMN1"
     CDS             5013..5039
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             5043..5069
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             5073..5099
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     misc_feature    5114..5702
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    5768..5816
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      6018..6446
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        6460..6789
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             6856..7650
                     /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    7826..7947
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(7996..8012)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    8020..8036
                     /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(8044..8074)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    8089..8110
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(8398..8986)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(9157..10017)
                     /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(10018..10122)
                     /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 ccaccatgga aggcactcat tgtaccctcc
      781 aattgcataa gcccattacg gaactctgct acatcagctt ctgtcttcca aagggggaag
      841 tcagaggatt ttcatacaag ggcactgtaa ctctagacag atccaataaa ggttttcata
      901 actgctacca agtcagggag gagtcagaca tcatcagcct cagccaggag ccggacgaac
      961 atccaggcga catatttttc aagcagactc ccacgaaaga cattctaact gagctgtaca
     1021 aactcacaac agagagggag agactgctaa ccaatctcct gagctcagac cacatcctgg
     1081 ggatcacgat ggggaaccag gaggggaagc tgcaagagct gtccgtgagc ctggcccccg
     1141 aggatgactg tttccagagt gctggtgact ggcagggaga gctccccgtg ggccctctca
     1201 ataagaggag cacccacggg aacaaaaagc ctcggaggtc tagtggaagg agagagagct
     1261 ttggggccct tccacagaag aggaccaaaa gaaaagggcg tggaggccga gaatcagctc
     1321 ctctgatggg caaggacaag atctgttcca gccactccct tcctctttct agaacaaggc
     1381 ctaacctttg ggtactagag gagaaaggaa atctgctccc gaatggggca cttgcctgct
     1441 ccctgcagag gagagagagc tgccccccag atattcccaa gacgccagac acagaccttg
     1501 gctttgggag ctttgagacg gctttcaagg acactgggct tggaagagaa gtgctgcccc
     1561 ctgactgcag ctccacagag gcaggagggg atggcattcg gaggccgccg agcgggctgg
     1621 agcatcagca aacaggtttg tctgaaagtc accaggaccc tgagaagcat ccagaggcag
     1681 aaaaggatga gatggagaag ccggctaagc ggacttgcaa gcagaaacct gtctccaaag
     1741 tggtggccaa agttcaggac ctgtcctccc aggtacaaag agtagttaaa acgcattcta
     1801 agggtaagga gacgattgcc attcgcccag cagcccacgc tgagtttgta cccaaagccg
     1861 acttgctcac cctcccggga gctgaggctg gggctcatgg ctccaggcgg cagggcaagg
     1921 agcggcaagg ggataggtca tcgcagtcgc cagccgggga aacagcctcc atttctagtg
     1981 tgtcggccag tgccgagggg gccgtgaaca aggtccccct gaaggtgata gagagtgaga
     2041 agttagatga agcccctgag gggaaaagac tgggcttccc tgtccacacg agtgtccctc
     2101 acactcgccc agaaacgaga aacaagagga gagccgggtt gccccttggt ggccacaagt
     2161 ccttgtttct ggatctgccc cacaaagtag gtcctgactc ctcacaaccc agaggtgata
     2221 agaagaagcc atccccacca gcaccggcag ctcttggcaa ggtgtttaat aattcagcct
     2281 cgcagtccag cacacacaaa cagacgtcac ctgttccctc gcctctgtct ccaaggctcc
     2341 ccagccctca gcagcatcac aggatcctcc ggctccctgc attgcctggt gagagggaag
     2401 ctgctcttaa tgactctcct tgtagaaaga gccgtgtctt ctctgggtgc gtctctgctg
     2461 acaccttgga gccaccatcc tctgcaaagg tcacggagac caaaggagcc agcccggcct
     2521 tcctcagagc aggccaacct cggttggtgc ctggggaaac tttggaaaag agcttggggc
     2581 cagggaagac cacagctgag ccccagcacc agtcacctcc aggtatctcc tctgagggct
     2641 ttccctggga cggcttcaat gagcagacac ctaaagacct tcccaacaga gatggaggcg
     2701 cgtgggttct gggctacaga gcgggaccag cctgtccatt tttgcttcat gaggaaaggg
     2761 agaagtcaaa caggagcgaa ttgtacttgg atctccatcc tgaccacagc ctgactgagc
     2821 aggatgacag gactcctggc agacttcaag ctgtctggcc acccccaaag acaaaagaca
     2881 cagaagaaaa agtgggactg aagtacactg aagcagaata ccaagctgct attttacact
     2941 tgaagaggga gcacaaagaa gaaattgaaa acctgcaggc acagtttgaa cttcgggcat
     3001 ttcatatccg gggcgagcat gcaatgataa cagcgagact agaagaaacc attgaaaatc
     3061 tgaaacacga gctagaacac agatggcgag ggggttgtga agagaggaaa gatgtgtgca
     3121 tttccaccga tgatgactgc cctccaaaga ccttcagaaa tgtgtgcgtc cagacagaca
     3181 gagagacctt cctcaagccc tgtgaaagtg aaagcaagac aaccagaagt aatcaattag
     3241 tacccaaaaa gctgaatatc tcctctttaa gccagctctc acccccaaat gaccacaaag
     3301 acatccatgc agcactccag ccaatggagg gcatggcatc aaatcagcag aaggcattgc
     3361 ctccgcctcc cgcatccatc cctccccctc cgcccctccc ctcaggactt ggatctttgt
     3421 ctcccgcacc tccaatgcca cctgtgagtg ctgggccacc gctaccacct ccgcctcctc
     3481 caccgccacc tctacctcca ccttcaagtg ctggacctcc accaccccca cccccacccc
     3541 cacttcctaa ctcccctgcc ccacccaacc ctggagggcc tcctcctgct ccaccacccc
     3601 caggacttgc acccccacct ccccctggac tcttctttgg acttggctct tcttccagtc
     3661 aatgtcctcg aaaaccagcc atcgagccca gttgtcccat gaagccttta tattggacta
     3721 ggatacaaat aagtgatagg agccaaaatg ctacaccaac cttatgggac tccttagaag
     3781 aacctgacat tcgggacccc agtgaatttg agtatttatt ctccaaagac acaactcaac
     3841 agaagaaaaa acctctgtca gagacttatg agaagaaaaa caaggtcaaa aagatcatca
     3901 aattgttgga tggaaaacga tctcaaactg tgggaatctt gatatctagt ttacatttag
     3961 aaatgaagga tatccaacag gccattttca atgtggatga ctccgtggtt gatctggaga
     4021 ccctggcagc cttatatgaa aacagagccc aagaggatga gctggttaaa ataagaaagt
     4081 attacgagac atccaaagaa gaagaactga agctgctgga taaacctgag caatttttac
     4141 atgagttagc ccagattcct aattttgctg aacgtgccca gtgcataatc ttcagatctg
     4201 tcttttctga gggtatcacc tccttgcaca gaaaggtaga gatcatcacg cgagcttcta
     4261 aggacttgct gcacgtgaag agcgtgaagg atattttagc tctcatcttg gcttttggaa
     4321 attatatgaa tggaggaaat aggactcggg gacaagccga tggatatagc ttagaaattc
     4381 tgcccaaact caaggatgtc aaaagtcggg ataatgggat taatctggtg gactacgttg
     4441 ttaagtatta cctgcgttac tatgatcagg aagctggaac agaaaagagt gttttcccct
     4501 tgccggaacc acaggatttc tttctggcct cccaagtcaa gtttgaagac ctcataaaag
     4561 atttgagaaa actgaagagg caactagaag caagtgagaa acagatggtg gtggtgtgca
     4621 aggagtcccc aaaggagtat ctccagcctt tcaaggacaa actagaggag ttcttccaaa
     4681 aagccaaaaa agagcataag atggaagaaa gtcacttgga gaatgcacag aaaagttttg
     4741 aaacaacagt acgatatttt gggatgaagc caaagtctgg tgagaaggag atcacaccca
     4801 gctacgtgtt tatggtgtgg tatgagttct gcagtgactt caagacaatt tggaaacggg
     4861 agagtaaaaa catatctaaa gaaagattga aaatggctca ggaatcagtc agcaagttga
     4921 cttcagagaa gaaagtggag acaaagaaaa tcaatcccac tgctagcctg aaagaaagac
     4981 tgcgtcagaa ggaagccagt gtgaccacta actacccata cgacgtacct gattatgcaa
     5041 cttatcctta tgatgtacct gattatgcta gctatcctta tgatgtccca gactacgcat
     5101 aggatatctc gacaatcaac ctctggatta caaaatttgt gaaagattga ctggtattct
     5161 taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc
     5221 tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct
     5281 ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga
     5341 cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc
     5401 tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac
     5461 aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaagc tgacgtcctt
     5521 tccatggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt
     5581 cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc
     5641 tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc
     5701 gcctggaaac gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc
     5761 gctatgacgg caataaaaag acagaataaa acgcacgggt gttgggtcgt ttgttcataa
     5821 acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg
     5881 ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc
     5941 agggctcgca gccaacgtcg gggcggcagg ccctgccata gcagatctgc gcagctgggg
     6001 ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     6061 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     6121 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc ggggcatccc
     6181 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     6241 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     6301 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     6361 ctattctttt gatttataag ggattttggg gatttcggcc tattggttaa aaaatgagct
     6421 gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga
     6481 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     6541 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     6601 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     6661 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     6721 gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     6781 ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca agagacagga
     6841 tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg
     6901 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
     6961 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
     7021 gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
     7081 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
     7141 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
     7201 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
     7261 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag
     7321 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag
     7381 gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat
     7441 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg
     7501 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
     7561 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
     7621 ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgcg aaatgaccga
     7681 ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag
     7741 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
     7801 catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata
     7861 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     7921 tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag
     7981 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     8041 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     8101 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     8161 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     8221 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     8281 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     8341 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     8401 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     8461 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     8521 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     8581 ggcgctttct caatgctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     8641 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     8701 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     8761 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     8821 ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     8881 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     8941 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     9001 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     9061 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     9121 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     9181 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     9241 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     9301 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     9361 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     9421 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     9481 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     9541 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     9601 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     9661 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     9721 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     9781 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     9841 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     9901 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     9961 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
    10021 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
    10081 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
    10141 gccacctgac gtcgacggat cgggagatct cccgatcccc tatggtcgac tctcagtaca
    10201 atctgctctg atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc
    10261 gctgagtagt gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc
    10321 atgaagaatc tgcttagg
//
