LOCUS       V035575                 6807 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V035575
VERSION     V035575
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        235..614
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        615..818
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        863..881
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     regulatory      895..904
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             904..927
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     CDS             946..2295
                     /label="Irf4(NM_013674)"
                     /note="Irf4(NM_013674)"
                     /gene="Irf4"
     promoter        complement(2359..2377)
                     /note="promoter for bacteriophage SP6 RNA polymerase"
                     /label="SP6 promoter"
     polyA_signal    2403..2627
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2673..3101
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3115..3445
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3512..4306
                     /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    4480..4601
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4650..4666)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4674..4690
                     /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(4698..4728)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4743..4764
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5052..5640)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5811..6671)
                     /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(6672..6776)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggac tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttgccacc
      901 atggactaca aggacgacga tgataagggt accgagctcg gatccatgaa cttggagacg
      961 ggcagccggg gctcagagtt cggcatgagc gcagtgagct gcggcaatgg gaaactccga
     1021 cagtggttga tcgaccagat cgacagcggc aagtaccccg ggctggtgtg ggagaacgag
     1081 gagaagagcg tcttccgcat cccgtggaaa cacgcgggca agcaggacta caatcgtgag
     1141 gaggacgctg ccctcttcaa ggcttgggca ttgtttaaag gcaagttccg agaagggatc
     1201 gacaagccag atcctcctac ttggaagaca agattacgat gtgctctgaa caagagcaat
     1261 gactttgagg aattggtcga gaggagccag ctggatatct ctgacccata caaggtgtac
     1321 aggattgttc cagagggagc caaaaaagga gcaaagcagc tcactttgga tgacacacag
     1381 atggccatgg gccaccccta ccccatgaca gcaccttatg gctctctgcc agcccagcag
     1441 gttcataact acatgatgcc accccatgac aggagctgga gggattatgc ccctgaccag
     1501 tcacacccag aaatcccata tcaatgtcct gtgacgtttg gcccacgagg ccaccactgg
     1561 caaggcccat cttgtgaaaa tggttgccag gtgacaggaa ccttttatgc ttgtgcccca
     1621 cctgagtccc aggctcctgg aatccccatt gagccaagca taaggtctgc tgaagccttg
     1681 gcgctctcag actgccggct gcatatctgc ctgtattacc gggacatcct cgtgaaagag
     1741 ctgaccacga cgagccctga aggctgccgg atctcccacg gacacaccta tgatgttagc
     1801 aacctggacc aggtcctgtt tccctacccg gacgacaatg gacagaggaa gaacattgag
     1861 aagttgctga gccacctgga gaggggactg gtcctctgga tggctccaga tgggctttat
     1921 gccaaaagac tctgccagag taggatctac tgggatgggc ccctggcact gtgcagcgat
     1981 cggcccaaca agctagaaag agaccagact tgcaagctct ttgacacaca gcagtttcta
     2041 tcagagctgc aagtgtttgc tcaccatggc cggccagcac cgagattcca ggtgactctg
     2101 tgctttggtg aggagtttcc agaccctcag agacagagga agctcatcac agctcatgtg
     2161 gaacctctgc tagccagaca actgtattac tttgctcaac aaaacactgg acatttcctg
     2221 aggggctacg agttacctga acacgttacc actccagatt accaccgctc cctccgtcat
     2281 tcttccatcc aagagtgaga attctgcaga tatccatcac actggcggcc gctcgagcat
     2341 gcatctagag ggccctattc tatagtgtca cctaaatgct agagctcgct gatcagcctc
     2401 gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac
     2461 cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg
     2521 tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga
     2581 ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga
     2641 aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc
     2701 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     2761 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     2821 aaatcggggc atccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     2881 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     2941 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     3001 caaccctatc tcggtctatt cttttgattt ataagggatt ttggggattt cggcctattg
     3061 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt
     3121 cagttagggt gtggaaagtc cccaggctcc ccaggcaggc agaagtatgc aaagcatgca
     3181 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     3241 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
     3301 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     3361 ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt
     3421 ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc
     3481 tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg
     3541 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg
     3601 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa
     3661 gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct
     3721 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga
     3781 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc
     3841 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac
     3901 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc
     3961 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact
     4021 gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga
     4081 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg
     4141 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga
     4201 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga
     4261 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg
     4321 ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc
     4381 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc
     4441 cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat
     4501 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     4561 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg
     4621 acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     4681 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     4741 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     4801 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     4861 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     4921 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     4981 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     5041 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     5101 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     5161 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     5221 ccttcgggaa gcgtggcgct ttctcaatgc tcacgctgta ggtatctcag ttcggtgtag
     5281 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     5341 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     5401 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     5461 aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg
     5521 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     5581 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     5641 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     5701 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     5761 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     5821 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     5881 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     5941 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     6001 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     6061 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     6121 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     6181 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     6241 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     6301 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     6361 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     6421 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     6481 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     6541 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     6601 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     6661 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     6721 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     6781 tttccccgaa aagtgccacc tgacgtc
//
