LOCUS       V020754                 6989 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V020754
VERSION     V020754
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..1669
                     /label="Irf1(NM_008390)"
                     /note="Irf1(NM_008390)"
                     /gene="Irf1"
     CDS             1676..1741
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    1765..2353
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2419..2467
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      2669..3097
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3111..3440
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3507..4301
                     /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    4477..4598
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4647..4663)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4671..4687
                     /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(4695..4725)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4740..4761
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5049..5637)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5808..6668)
                     /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(6669..6773)
                     /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 ccatgccaat cactcgaatg cggatgagac cctggctaga
      721 gatgcagatt aattccaacc aaatcccagg gctgatctgg atcaataaag aagagatgat
      781 cttccagatt ccatggaagc acgctgctaa gcacggctgg gacatcaaca aggatgcctg
      841 tctgttccgg agctgggcca ttcacacagg ccgatacaaa gcaggagaaa aagagccaga
      901 tcccaagaca tggaaggcaa acttccgttg tgccatgaac tccctgccag acatcgagga
      961 agtgaaggat cagagtagga acaagggcag ctctgctgtg cgggtgtacc ggatgctgcc
     1021 acccctcacc aggaaccaga ggaaagagag aaagtccaag tccagccgag acactaagag
     1081 caaaaccaag aggaagctgt gtggagatgt tagcccggac actttctctg atggactcag
     1141 cagctctacc ctacctgatg accacagcag ttacaccact cagggctacc tgggtcagga
     1201 cttggatatg gaaagggaca taactccagc actgtcaccg tgtgtcgtca gcagcagtct
     1261 ctctgagtgg catatgcaga tggacattat accagatagc accactgatc tgtataacct
     1321 acaggtgtca cccatgcctt ccacctccga agccgcaaca gacgaggatg aggaagggaa
     1381 gatagccgaa gaccttatga agctctttga acagtctgag tggcagccga cacacatcga
     1441 tggcaaggga tacttgctca atgagccagg gacccagctc tcttctgtct atggagactt
     1501 cagctgcaaa gaggaaccag agattgacag ccctcgaggg gacattggga taggcataca
     1561 acatgtcttc acggagatga agaatatgga ctccatcatg tggatggaca gcctgctggg
     1621 caactctgtg aggctgccgc cctctattca ggccattcct tgtgcaccac tcgaggacta
     1681 caaagaccat gacggtgatt ataaagatca tgacatcgac tacaaggatg acgatgacaa
     1741 gtagtgaggg cccgatatct cgacaatcaa cctctggatt acaaaatttg tgaaagattg
     1801 actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct
     1861 ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg
     1921 ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact
     1981 gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc
     2041 gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc
     2101 cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaag
     2161 ctgacgtcct ttccatggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc
     2221 ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg
     2281 gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg
     2341 gccgcctccc cgcctggaaa cgggggaggc taactgaaac acggaaggag acaataccgg
     2401 aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggg tgttgggtcg
     2461 tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga
     2521 ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg
     2581 ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat agcagatctg
     2641 cgcagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg
     2701 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     2761 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     2821 cggggcatcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     2881 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     2941 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     3001 cctatctcgg tctattcttt tgatttataa gggattttgg ggatttcggc ctattggtta
     3061 aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt
     3121 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     3181 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     3241 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     3301 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     3361 cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg
     3421 gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc
     3481 aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc
     3541 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct
     3601 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg
     3661 acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca
     3721 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc
     3781 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga
     3841 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc
     3901 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc
     3961 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg
     4021 ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct
     4081 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc
     4141 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc
     4201 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc
     4261 agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcgc
     4321 gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc
     4381 ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag
     4441 cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat
     4501 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     4561 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc
     4621 tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     4681 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     4741 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     4801 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     4861 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     4921 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     4981 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     5041 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     5101 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     5161 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     5221 tcgggaagcg tggcgctttc tcaatgctca cgctgtaggt atctcagttc ggtgtaggtc
     5281 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     5341 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     5401 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     5461 tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag
     5521 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     5581 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     5641 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     5701 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     5761 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     5821 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     5881 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     5941 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     6001 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     6061 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     6121 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     6181 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     6241 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     6301 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     6361 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     6421 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     6481 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     6541 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     6601 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     6661 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     6721 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     6781 ccccgaaaag tgccacctga cgtcgacgga tcgggagatc tcccgatccc ctatggtcga
     6841 ctctcagtac aatctgctct gatgccgcat agttaagcca gtatctgctc cctgcttgtg
     6901 tgttggaggt cgctgagtag tgcgcgagca aaatttaagc tacaacaagg caaggcttga
     6961 ccgacaattg catgaagaat ctgcttagg
//
