LOCUS       V030690                 8162 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V030690
VERSION     V030690
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             683..2821
                     /label="MIPEP(NM_005932)"
                     /note="MIPEP(NM_005932)"
                     /gene="MIPEP"
     CDS             2828..2854
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             2858..2884
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             2888..2914
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     misc_feature    2938..3526
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3592..3640
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3842..4270
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4284..4613
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4680..5474
                     /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    5650..5771
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5820..5836)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5844..5860
                     /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(5868..5898)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5913..5934
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6222..6810)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6981..7841)
                     /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(7842..7946)
                     /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 ggatccgcca ccatgctgtg cgtcggaagg ctgggcggct tgggagccag
      721 agcagcagct ctgccgcccc gccgggcggg ccggggaagc ctcgaagccg ggatccgggc
      781 ccgaagggtc agcaccagct ggtctcccgt gggcgccgcc ttcaatgtca agccccaggg
      841 cagccgcttg gacctgttcg gcgagcgccg gggtcttttt ggagttcctg agctgagtgc
      901 cccagaagga tttcatattg cacaagaaaa agccttgaga aagacagaat tgcttgtgga
      961 ccgtgcatgt tccaccccac ctgggcccca gaccgtgctg atcttcgatg agctctcgga
     1021 ttccttatgc agagtggccg acttggctga ttttgtgaaa atcgctcacc ctgagccagc
     1081 attcagagaa gctgcggaag aagcttgtag aagtattggc accatggtag agaagttgaa
     1141 cacaaatgtg gatttatatc aaagtttgca aaaattacta gctgataaaa aacttgtgga
     1201 ttcccttgat ccagaaacaa ggcgagtggc tgaactgttt atgtttgatt ttgaaattag
     1261 tggaatccat ctagacaaag aaaagcgtaa aagagcagtg gacctcaatg ttaaaatctt
     1321 ggatttgagt agtacatttc ttatgggaac caattttccc aacaagattg agaagcatct
     1381 cttaccagaa cacattcgtc gtaactttac atctgctggg gatcatatca taattgatgg
     1441 tctccacgca gaatcaccag atgacttggt gcgagaagct gcttataaaa tttttcttta
     1501 tcccaatgct ggtcaattga aatgtttaga agaattgctc agcagcagag atcttctggc
     1561 aaagttggtg gggtattcca cgttttctca cagggctctc caaggaacga tagctaaaaa
     1621 tccagagact gtcatgcagt tccttgaaaa actatctgac aaactttctg aaagaactct
     1681 gaaagatttt gagatgatac gagggatgaa aatgaaactg aatcctcaaa attccgaagt
     1741 aatgccctgg gacccccctt actacagtgg tgtgattcgt gcagaaaggt ataatattga
     1801 gcccagccta tattgcccgt ttttctctct tggagcatgc atggaaggcc tgaatatttt
     1861 gcttaacaga ctgttgggga tttcattata tgcagagcag cctgcaaaag gagaggtgtg
     1921 gagcgaagat gtccgaaaac tggctgttgt tcatgaatct gaaggattgt tggggtacat
     1981 ttactgtgat ttttttcagc gagcagacaa accacatcag gattgccatt tcactatccg
     2041 tggaggcaga ctaaaggaag atggagacta tcaactccca gttgtagttc ttatgctgaa
     2101 tcttccccgt tcctcaagga gttctccaac tttgctaact cctagcatga tggaaaatct
     2161 tttccatgaa atgggacatg ccatgcattc aatgctagga cgtactcgtt accaacacgt
     2221 cactgggacc aggtgcccta ctgattttgc tgaggttcct tctattctga tggagtactt
     2281 tgcaaatgat tatcgagtag ttaaccaatt tgccagacat tatcagactg gacagccact
     2341 gccaaaaaat atggtgtctc gtctttgtga atctaaaaag gtttgtgctg cagctgatat
     2401 gcaacttcag gtcttttatg ccactctgga tcaaatctac catgggaagc atcccctgag
     2461 gaattcaacc acagacattc tcaaggaaac acaagagaaa ttctatggcc taccatatgt
     2521 tccaaatact gcctggcagc tgcgattcag ccacctcgtg gggtatggtg ctagatatta
     2581 ctcttacctc atgtccagag cggtcgcctc catggtttgg aaggagtgtt ttctacagga
     2641 tcctttcaac agggctgccg gggagcgcta tcgcagggag atgctggccc acggtggagg
     2701 cagggagccc atgctcatgg ttgaaggtat gcttcagaag tgtccttctg ttgatgactt
     2761 cgtaagtgcc ctcgtttccg acttggatct ggacttcgaa actttcctca tggattctga
     2821 actcgagtac ccatacgacg tacctgatta tgcaacttat ccttatgatg tacctgatta
     2881 tgctagctat ccttatgatg tcccagacta cgcatagtga gggcccgata tctcgacaat
     2941 caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct
     3001 tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg
     3061 gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg
     3121 cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt
     3181 tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt
     3241 gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg
     3301 ggcactgaca attccgtggt gttgtcgggg aagctgacgt cctttccatg gctgctcgcc
     3361 tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat
     3421 ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc
     3481 cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcctgg aaacggggga
     3541 ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa
     3601 aaagacagaa taaaacgcac gggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc
     3661 cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata cgcccgcgtt
     3721 tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct cgcagccaac
     3781 gtcggggcgg caggccctgc catagcagat ctgcgcagct ggggctctag ggggtatccc
     3841 cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     3901 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     3961 acgttcgccg gctttccccg tcaagctcta aatcggggca tccctttagg gttccgattt
     4021 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     4081 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     4141 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     4201 taagggattt tggggatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     4261 aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
     4321 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
     4381 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     4441 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     4501 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg
     4561 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc
     4621 cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga tcgtttcgca
     4681 tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg
     4741 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag
     4801 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc
     4861 aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc
     4921 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg
     4981 atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc
     5041 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca
     5101 tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag
     5161 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg
     5221 gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg
     5281 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca
     5341 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc
     5401 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg
     5461 acgagttctt ctgagcggga ctctggggtt cgcgaaatga ccgaccaagc gacgcccaac
     5521 ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc
     5581 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc
     5641 gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     5701 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     5761 aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg
     5821 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     5881 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     5941 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     6001 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     6061 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     6121 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     6181 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     6241 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     6301 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     6361 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcaatgc
     6421 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     6481 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     6541 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     6601 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     6661 aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     6721 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     6781 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     6841 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     6901 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     6961 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     7021 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     7081 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     7141 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     7201 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     7261 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     7321 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     7381 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     7441 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     7501 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     7561 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     7621 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     7681 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     7741 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     7801 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     7861 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     7921 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac
     7981 ggatcgggag atctcccgat cccctatggt cgactctcag tacaatctgc tctgatgccg
     8041 catagttaag ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga
     8101 gcaaaattta agctacaaca aggcaaggct tgaccgacaa ttgcatgaag aatctgctta
     8161 gg
//
