LOCUS       V040250                 8286 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V040250
VERSION     V040250
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..2966
                     /label="DDX50(NM_024045)"
                     /note="DDX50(NM_024045)"
                     /gene="DDX50"
     CDS             2973..3038
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3062..3650
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3716..3764
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3966..4394
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4408..4737
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4804..5598
                     /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    5774..5895
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5944..5960)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5968..5984
                     /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(5992..6022)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6037..6058
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6346..6934)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7105..7965)
                     /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(7966..8070)
                     /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 ccaccatgcc tgggaaactc ctctgggggg
      781 acattatgga gctggaagca cccttggagg agtccgagag ccagaagaag gagaggcaaa
      841 agagtgacag aaggaagtca aggcaccatt atgactcgga tgagaaatca gaaacaagag
      901 aaaatggtgt tacagatgac ctggatgctc ccaaggccaa aaaatctaaa atgaaagaga
      961 agctaaatgg agacactgaa gaaggattta atagactttc agatgaattc tccaaatctc
     1021 ataagtcaag aagaaaagat ctaccaaatg gagatataga tgaatatgaa aaaaaatcaa
     1081 agcgagtatc atctttagat acttctactc ataaatcaag tgataataaa ctagaggaga
     1141 ccttaacacg tgaacagaaa gaaggagcct tctccaattt tcctatttct gaagagacta
     1201 taaagcttct gaaaggtcga ggggtaacat atctctttcc tattcaagtt aagacctttg
     1261 gtcctgtata tgaaggaaaa gatttaatag ctcaagcacg gacaggaaca ggaaagacat
     1321 tctcttttgc catcccctta attgaaagac tccaaagaaa tcaagaaaca attaaaaaaa
     1381 gccgctcacc aaaggtactt gttttggctc caacaaggga actggcaaac caagtagcca
     1441 aagacttcaa agatataact aggaaactca gcgtggcgtg tttttatggt ggaacatcat
     1501 atcaaagcca aattaatcat attcgaaatg gtattgacat cttggttgga acacctggtc
     1561 gtatcaaaga ccatctgcag agtggccgat tggatctttc taaactgcga catgttgtgc
     1621 ttgatgaagt ggatcagatg ttagatttag gtttcgctga acaagttgaa gatattattc
     1681 atgaatccta caaaactgat tctgaagaca atcctcagac tttacttttt tctgcaactt
     1741 gcccacagtg ggtatacaaa gttgcaaaaa aatacatgaa atccagatat gaacaggttg
     1801 accttgttgg aaaaatgact caaaaggctg caactactgt ggaacatttg gccatccagt
     1861 gtcattggtc tcagaggcca gcagttattg gagatgtcct tcaagtctac agtgggtctg
     1921 aagggagggc tattattttc tgtgagacca agaagaatgt aactgaaatg gccatgaatc
     1981 cacacataaa acagaatgcc cagtgtttac atggggacat tgcacagtca caaagagaaa
     2041 ttacactaaa aggcttcaga gaaggtagtt ttaaagtttt ggtggcaacc aatgtggctg
     2101 cccgtggttt ggacattcct gaagttgacc tggtgattca aagttctcct cctcaggatg
     2161 ttgagtccta tatccatcgc tctggacgca caggtagagc tggacggaca gggatttgta
     2221 tatgttttta tcaaccaaga gaaagaggtc aactaagata tgtggaacaa aaagcaggaa
     2281 ttacttttaa acgtgtaggt gttccttcta caatggattt agttaaatct aaaagcatgg
     2341 atgccatcag gtctctggct tccgtttctt acgctgctgt tgattttttc cgaccatcag
     2401 ctcagagact gatagaagag aaaggtgcag tggatgcatt ggctgcagct ttagcccaca
     2461 tttctggtgc atcaagcttt gaaccacgat ctttgatcac ctctgataag gggtttgtga
     2521 ccatgactct ggaaagccta gaggaaatac aggatgtcag ctgtgcttgg aaagaactta
     2581 acagaaagct gagtagtaat gcagtgtctc agattaccag aatgtgcctc ctgaaaggaa
     2641 atatgggtgt ttgctttgat gttcctacaa ctgagtcaga aaggttacag gcagagtggc
     2701 atgattccga ctggatactc tcagtgccag ccaaattacc tgaaattgaa gaatattatg
     2761 atggaaacac atcttctaat tccagacaga ggagtggctg gtcaagtggt cgatcaggcc
     2821 ggtcaggccg gtcaggtggt cgatctggcg gccggtcagg tagacagagt cgacaaggaa
     2881 gtcgctcagg aagtcgacaa gatggtagaa gacgaagtgg gaatagaaat cgatcaagaa
     2941 gtgggggcca caaacggagt tttgacggat ccgactacaa agaccatgac ggtgattata
     3001 aagatcatga catcgactac aaggatgacg atgacaagta gtgagggccc gatatctcga
     3061 caatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc
     3121 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg
     3181 tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt
     3241 gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac
     3301 tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc
     3361 tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct
     3421 gttgggcact gacaattccg tggtgttgtc ggggaagctg acgtcctttc catggctgct
     3481 cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct
     3541 caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct
     3601 tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc ctggaaacgg
     3661 gggaggctaa ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca
     3721 ataaaaagac agaataaaac gcacgggtgt tgggtcgttt gttcataaac gcggggttcg
     3781 gtcccagggc tggcactctg tcgatacccc accgagaccc cattggggcc aatacgcccg
     3841 cgtttcttcc ttttccccac cccacccccc aagttcgggt gaaggcccag ggctcgcagc
     3901 caacgtcggg gcggcaggcc ctgccatagc agatctgcgc agctggggct ctagggggta
     3961 tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     4021 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     4081 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg ggcatccctt tagggttccg
     4141 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
     4201 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     4261 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
     4321 tttataaggg attttgggga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     4381 atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc
     4441 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga
     4501 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     4561 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     4621 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc
     4681 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag
     4741 ctcccgggag cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt
     4801 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     4861 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
     4921 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     4981 ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     5041 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     5101 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     5161 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     5221 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     5281 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc
     5341 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     5401 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     5461 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     5521 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     5581 cttgacgagt tcttctgagc gggactctgg ggttcgcgaa atgaccgacc aagcgacgcc
     5641 caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg
     5701 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt
     5761 cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     5821 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     5881 catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc
     5941 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     6001 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     6061 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     6121 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     6181 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     6241 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     6301 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     6361 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     6421 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     6481 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     6541 atgctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     6601 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     6661 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     6721 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     6781 tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     6841 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     6901 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     6961 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     7021 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     7081 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     7141 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     7201 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     7261 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     7321 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     7381 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     7441 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     7501 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     7561 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     7621 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     7681 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     7741 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     7801 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     7861 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     7921 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     7981 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     8041 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     8101 cgacggatcg ggagatctcc cgatccccta tggtcgactc tcagtacaat ctgctctgat
     8161 gccgcatagt taagccagta tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc
     8221 gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg acaattgcat gaagaatctg
     8281 cttagg
//
