LOCUS       V033150                 7224 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V033150
VERSION     V033150
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..1904
                     /label="MICB(NM_005931)"
                     /note="MICB(NM_005931)"
                     /gene="MICB"
     CDS             1911..1976
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2000..2588
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2654..2702
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      2904..3332
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3346..3675
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3742..4536
                     /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    4712..4833
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4882..4898)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4906..4922
                     /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(4930..4960)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4975..4996
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5284..5872)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6043..6903)
                     /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(6904..7008)
                     /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 ccaccatggg gctgggccgg gtcctgctgt
      781 ttctggccgt cgccttccct tttgcacccc cggcagccgc cgctgagccc cacagtcttc
      841 gttacaacct catggtgctg tcccaggatg gatctgtgca gtcagggttt ctcgctgagg
      901 gacatctgga tggtcagccc ttcctgcgct atgacaggca gaaacgcagg gcaaagcccc
      961 agggacagtg ggcagaaaat gtcctgggag ctaagacctg ggacacagag accgaggact
     1021 tgacagagaa tgggcaagac ctcaggagga ccctgactca tatcaaggac cagaaaggag
     1081 gcttgcattc cctccaggag attagggtct gtgagatcca tgaagacagc agcaccaggg
     1141 gctcccggca tttctactac gatggggagc tcttcctctc ccaaaacctg gagactcaag
     1201 aatcgacagt gccccagtcc tccagagctc agaccttggc tatgaacgtc acaaatttct
     1261 ggaaggaaga tgccatgaag accaagacac actatcgcgc tatgcaggca gactgcctgc
     1321 agaaactaca gcgatatctg aaatccgggg tggccatcag gagaacagtg ccccccatgg
     1381 tgaatgtcac ctgcagcgag gtctcagagg gcaacatcac cgtgacatgc agggcttcca
     1441 gcttctatcc ccggaatatc acactgacct ggcgtcagga tggggtatct ttgagccaca
     1501 acacccagca gtggggggat gtcctgcctg atgggaatgg aacctaccag acctgggtgg
     1561 ccaccaggat tcgccaagga gaggagcaga ggttcacctg ctacatggaa cacagcggga
     1621 atcacggcac tcaccctgtg ccctctggga aggcgctggt gcttcagagt caacggacag
     1681 actttccata tgtttctgct gctatgccat gttttgttat tattattatt ctctgtgtcc
     1741 cttgttgcaa gaagaaaaca tcagcggcag agggtccaga gcttgtgagc ctgcaggtcc
     1801 tggatcaaca cccagttggg acaggagacc acagggatgc agcacagctg ggatttcagc
     1861 ctctgatgtc agctactggg tccactggtt ccactgaggg caccggatcc gactacaaag
     1921 accatgacgg tgattataaa gatcatgaca tcgactacaa ggatgacgat gacaagtagt
     1981 gagggcccga tatctcgaca atcaacctct ggattacaaa atttgtgaaa gattgactgg
     2041 tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta
     2101 tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct
     2161 gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt
     2221 tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac
     2281 tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg
     2341 ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaagctgac
     2401 gtcctttcca tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg
     2461 ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct
     2521 gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc
     2581 ctccccgcct ggaaacgggg gaggctaact gaaacacgga aggagacaat accggaagga
     2641 acccgcgcta tgacggcaat aaaaagacag aataaaacgc acgggtgttg ggtcgtttgt
     2701 tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca
     2761 ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga
     2821 aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcag atctgcgcag
     2881 ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     2941 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     3001 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     3061 catcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     3121 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     3181 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     3241 ctcggtctat tcttttgatt tataagggat tttggggatt tcggcctatt ggttaaaaaa
     3301 tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg
     3361 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     3421 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     3481 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     3541 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     3601 gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     3661 ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag
     3721 acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc
     3781 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat
     3841 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg
     3901 tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg
     3961 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta
     4021 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta
     4081 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc
     4141 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc
     4201 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg
     4261 ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg
     4321 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
     4381 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc
     4441 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
     4501 atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgcgaaat
     4561 gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta
     4621 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
     4681 ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta
     4741 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     4801 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag
     4861 ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     4921 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     4981 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     5041 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     5101 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     5161 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     5221 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     5281 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     5341 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     5401 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     5461 aagcgtggcg ctttctcaat gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     5521 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     5581 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     5641 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     5701 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt
     5761 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     5821 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     5881 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     5941 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     6001 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     6061 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     6121 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     6181 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     6241 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     6301 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     6361 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     6421 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     6481 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     6541 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     6601 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     6661 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     6721 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     6781 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     6841 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     6901 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     6961 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     7021 aaaagtgcca cctgacgtcg acggatcggg agatctcccg atcccctatg gtcgactctc
     7081 agtacaatct gctctgatgc cgcatagtta agccagtatc tgctccctgc ttgtgtgttg
     7141 gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa caaggcaagg cttgaccgac
     7201 aattgcatga agaatctgct tagg
//
