LOCUS       V021004                 8384 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V021004
VERSION     V021004
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..3007
                     /label="QARS1(NM_005051)"
                     /note="QARS1(NM_005051)"
                     /gene="QARS1"
     CDS             3047..3136
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     misc_feature    3160..3748
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3814..3862
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4064..4492
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4506..4835
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4902..5696
                     /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    5872..5993
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6042..6058)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6066..6082
                     /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(6090..6120)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6135..6156
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6444..7032)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7203..8063)
                     /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(8064..8168)
                     /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 ccatggcggc tctagactcc ctgtcgctct tcactagcct
      721 cggcctgagc gagcagaagg cccgcgagac gctcaagaac tcggctctga gcgcgcagct
      781 gcgcgaggcc gctactcagg ctcagcagac cctgggttcc accattgaca aagctaccgg
      841 gatcctgtta tatggcttgg cctcccgact cagggatacc cggcgtctct ccttccttgt
      901 aagctacata gccagtaaga agatccacac tgagccccag ctaagcgctg cccttgagta
      961 tgtgcggagt caccccttgg accccatcga cactgtggac ttcgagcggg aatgtggcgt
     1021 gggtgtcatt gtgaccccag agcagattga ggaggctgtg gaggctgcta ttaacaggca
     1081 ccggccccag ctcctggtgg aacgttacca tttcaacatg gggctgctga tgggagaggc
     1141 tcgggctgtg ctgaagtggg cagatggcaa aatgatcaag aatgaagtgg acatgcaggt
     1201 cctccacctt ctgggcccca agttggaggc tgatctggag aagaagttca aggtggcaaa
     1261 agctcggcta gaagaaacag accggaggac ggcaaaggat gtggtggaga atggcgagac
     1321 tgctgaccag accctgtctc tgatggagca gctccggggg gaggccctta agttccacaa
     1381 gcctggtgag aactacaaga ccccaggcta tgtggtcact ccacacacca tgaatctact
     1441 aaagcagcac ctggagatta ctggtgggca ggtacgtacc cggttcccgc cagaacccaa
     1501 tggaatcctg catattggac atgccaaagc catcaatttc aactttggct atgccaaggc
     1561 caacaatggc atctgttttc tgcgttttga tgacaccaac cctgagaagg aggaagcaaa
     1621 gttcttcacg gccatctgtg acatggtagc ctggctaggc tacacacctt acaaagtcac
     1681 atatgcgtct gactattttg accagctata tgcgtgggct gtggagctca tccgcagggg
     1741 tctggcttat gtgtgccacc agcgaggaga ggagctcaaa ggccataata ctctgccttc
     1801 accctggaga gaccgtccca tggaggagtc actgctgctc tttgaggcaa tgcgcaaggg
     1861 caagttttca gagggcgagg ccacactacg gatgaagctg gtgatggagg atggcaagat
     1921 ggaccctgta gcctatcgag tcaagtatac accacaccac cgcacagggg acaaatggtg
     1981 catctatccc acctacgact acacacactg cctctgtgac tccatcgagc acatcactca
     2041 ctcactctgc accaaggaat tccaggcccg acgctcttcc tacttctggc tttgcaatgc
     2101 actggacgtc tattgccctg tgcagtggga gtatggccgc ctcaacctgc actatgctgt
     2161 tgtctctaag aggaagatcc tccagcttgt agcaactggt gctgtgcggg actgggatga
     2221 cccacggctc tttacactca cggccctgcg acggcggggc ttcccacctg aggccatcaa
     2281 caacttctgt gcccgggtgg gagtgactgt ggcacaaacc acaatggagc cacatcttct
     2341 agaagcctgt gtgcgtgatg tgctgaatga cacagcccca cgagccatgg ctgtgctgga
     2401 gtcactacgg gtcatcatca ccaactttcc tgctgccaag tccttggaca tccaggtgcc
     2461 caacttccca gctgatgaga ccaaaggctt ccatcaggtt ccctttgcac ccattgtctt
     2521 cattgagagg actgacttca aggaggagcc agagccagga tttaagcgcc tggcttgggg
     2581 ccagcctgtg ggcctgaggc atacaggcta cgtcattgag ctgcagcatg ttgtcaaggg
     2641 ccccagtggt tgtgtagaga gtctggaggt gacctgcaga cgggcagatg ctggagagaa
     2701 gccaaaggcc tttattcact gggtgtcaca gcctttgatg tgtgaggttc gcctctatga
     2761 gcgactattc cagcacaaga accctgaaga tcctactgag gtgcctggtg gatttttaag
     2821 tgacctgaac ctggcatcac tacacgtggt ggatgcagca ttagtggact gctctgtggc
     2881 cctggcaaaa cccttcgaca agttccagtt tgagcgtctt ggatatttct ccgtggatcc
     2941 agacagccat cagggaaagc ttgtctttaa ccgaactgtc acactgaagg aagacccagg
     3001 aaaggtggaa ttctgcagat atccagcaca gtggcggccg ctcgaggaac aaaaactaat
     3061 atcagaggaa gatttggaac agaaactaat cagtgaagaa gatttagagc aaaagttaat
     3121 ttccgaggag gacttatagt gagggcccga tatctcgaca atcaacctct ggattacaaa
     3181 atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac
     3241 gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc
     3301 ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt
     3361 ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc
     3421 tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc
     3481 gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg
     3541 gtgttgtcgg ggaagctgac gtcctttcca tggctgctcg cctgtgttgc cacctggatt
     3601 ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc
     3661 cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt
     3721 cggatctccc tttgggccgc ctccccgcct ggaaacgggg gaggctaact gaaacacgga
     3781 aggagacaat accggaagga acccgcgcta tgacggcaat aaaaagacag aataaaacgc
     3841 acgggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc
     3901 gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc
     3961 caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct
     4021 gccatagcag atctgcgcag ctggggctct agggggtatc cccacgcgcc ctgtagcggc
     4081 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     4141 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     4201 cgtcaagctc taaatcgggg catcccttta gggttccgat ttagtgcttt acggcacctc
     4261 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     4321 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     4381 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttggggatt
     4441 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt
     4501 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     4561 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
     4621 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
     4681 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     4741 ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt
     4801 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca
     4861 ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat
     4921 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac
     4981 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc
     5041 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc
     5101 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag
     5161 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc
     5221 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg
     5281 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc
     5341 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc
     5401 cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga
     5461 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
     5521 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg
     5581 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg
     5641 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg
     5701 gactctgggg ttcgcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc
     5761 gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc
     5821 tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt
     5881 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     5941 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
     6001 tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg
     6061 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     6121 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     6181 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     6241 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     6301 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     6361 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     6421 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     6481 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     6541 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     6601 tccgcctttc tcccttcggg aagcgtggcg ctttctcaat gctcacgctg taggtatctc
     6661 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     6721 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     6781 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     6841 acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc
     6901 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     6961 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     7021 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     7081 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     7141 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     7201 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     7261 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     7321 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     7381 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     7441 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     7501 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     7561 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     7621 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     7681 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     7741 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     7801 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     7861 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     7921 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     7981 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     8041 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     8101 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     8161 gttccgcgca catttccccg aaaagtgcca cctgacgtcg acggatcggg agatctcccg
     8221 atcccctatg gtcgactctc agtacaatct gctctgatgc cgcatagtta agccagtatc
     8281 tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa
     8341 caaggcaagg cttgaccgac aattgcatga agaatctgct tagg
//
