LOCUS       V024412                 7358 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V024412
VERSION     V024412
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"
     CDS             941..2848
                     /label="IL27RA(NM_004843)"
                     /note="IL27RA(NM_004843)"
                     /gene="IL27RA"
     CDS             2855..2920
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    2958..3182
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3228..3656
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3670..3999
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4066..4860
                     /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    5034..5155
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5204..5220)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5228..5244
                     /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(5252..5282)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5297..5318
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5606..6191)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6362..7222)
                     /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(7223..7327)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct 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 atgggtggag 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 gctggctagc
      901 gtttaaactt aagcttggta ccgagctcgg atccgccacc atgcggggag gcaggggcgc
      961 ccctttctgg ctgtggccgc tgcccaagct ggcgctgctg cctctgttgt gggtgctttt
     1021 ccagcggacg cgtccccagg gcagcgccgg gccactgcag tgctacggag ttggaccctt
     1081 gggcgacttg aactgctcgt gggagcctct tggggacctg ggagccccct ccgagttaca
     1141 cctccagagc caaaagtacc gttccaacaa aacccagact gtggcagtgg cagccggacg
     1201 gagctgggtg gccattcctc gggaacagct caccatgtct gacaaactcc ttgtctgggg
     1261 cactaaggca ggccagcctc tctggccccc cgtcttcgtg aacctagaaa cccaaatgaa
     1321 gccaaacgcc ccccggctgg gccctgacgt ggacttttcc gaggatgacc ccctggaggc
     1381 cactgtccat tgggccccac ctacatggcc atctcataaa gttctgatct gccagttcca
     1441 ctaccgaaga tgtcaggagg cggcctggac cctgctggaa ccggagctga agaccatacc
     1501 cctgacccct gttgagatcc aagatttgga gctagccact ggctacaaag tgtatggccg
     1561 ctgccggatg gagaaagaag aggatttgtg gggcgagtgg agccccattt tgtccttcca
     1621 gacaccgcct tctgctccaa aagatgtgtg ggtatcaggg aacctctgtg ggacgcctgg
     1681 aggagaggaa cctttgcttc tatggaaggc cccagggccc tgtgtgcagg tgagctacaa
     1741 agtctggttc tgggttggag gtcgtgagct gagtccagaa ggaattacct gctgctgctc
     1801 cctaattccc agtggggcgg agtgggccag ggtgtccgct gtcaacgcca caagctggga
     1861 gcctctcacc aacctctctt tggtctgctt ggattcagcc tctgcccccc gtagcgtggc
     1921 agtcagcagc atcgctggga gcacggagct actggtgacc tggcaaccgg ggcctgggga
     1981 accactggag catgtagtgg actgggctcg agatggggac cccctggaga aactcaactg
     2041 ggtccggctt ccccctggga acctcagtgc tctgttacca gggaatttca ctgtcggggt
     2101 cccctatcga atcactgtga ccgcagtctc tgcttcaggc ttggcctctg catcctccgt
     2161 ctgggggttc agggaggaat tagcacccct agtggggcca acgctttggc gactccaaga
     2221 tgcccctcca gggacccccg ccatagcgtg gggagaggtc ccaaggcacc agcttcgagg
     2281 ccacctcacc cactacacct tgtgtgcaca gagtggaacc agcccctccg tctgcatgaa
     2341 tgtgagtggc aacacacaga gtgtcaccct gcctgacctt ccttggggtc cctgtgagct
     2401 gtgggtgaca gcatctacca tcgctggaca gggccctcct ggtcccatcc tccggcttca
     2461 tctaccagat aacaccctga ggtggaaagt tctgccgggc atcctattct tgtggggctt
     2521 gttcctgttg gggtgtggcc tgagcctggc cacctctgga aggtgctacc acctaaggca
     2581 caaagtgctg ccccgctggg tctgggagaa agttcctgat cctgccaaca gcagttcagg
     2641 ccagccccac atggagcaag tacctgaggc ccagcccctt ggggacttgc ccatcctgga
     2701 agtggaggag atggagcccc cgccggttat ggagtcctcc cagcccgccc aggccaccgc
     2761 cccgcttgac tctgggtatg agaagcactt cctgcccaca cctgaggagc tgggccttct
     2821 ggggcccccc aggccacagg ttctggccct cgaggactac aaagaccatg acggtgatta
     2881 taaagatcat gacatcgact acaaggatga cgatgacaag tagtgagggc ccgtttaaac
     2941 ccgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc
     3001 cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga
     3061 aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga
     3121 cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat
     3181 ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag
     3241 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     3301 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     3361 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     3421 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     3481 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     3541 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
     3601 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt
     3661 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     3721 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     3781 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     3841 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     3901 ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag
     3961 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata
     4021 tccattttcg gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat
     4081 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     4141 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     4201 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg
     4261 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     4321 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     4381 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     4441 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     4501 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     4561 gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc
     4621 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     4681 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     4741 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     4801 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     4861 gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt
     4921 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
     4981 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt
     5041 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     5101 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     5161 tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg
     5221 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     5281 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     5341 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     5401 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     5461 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     5521 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     5581 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     5641 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     5701 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     5761 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     5821 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     5881 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     5941 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     6001 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     6061 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     6121 aacaaaccac cgctggtagc ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     6181 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     6241 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     6301 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     6361 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     6421 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     6481 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     6541 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     6601 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     6661 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     6721 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     6781 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     6841 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     6901 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     6961 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     7021 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     7081 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     7141 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     7201 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     7261 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     7321 ttccgcgcac atttccccga aaagtgccac ctgacgtc
//
