LOCUS       V021099                 8096 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V021099
VERSION     V021099
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..2743
                     /label="TGM2(NM_004613)"
                     /note="TGM2(NM_004613)"
                     /gene="TGM2"
     CDS             2783..2848
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2872..3460
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3526..3574
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3776..4204
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4218..4547
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4614..5408
                     /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    5584..5705
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5754..5770)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5778..5794
                     /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(5802..5832)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5847..5868
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6156..6744)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6915..7775)
                     /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(7776..7880)
                     /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 ccatggccga ggagctggtc ttagagaggt gtgatctgga
      721 gctggagacc aatggccgag accaccacac ggccgacctg tgccgggaga agctggtggt
      781 gcgacggggc cagcccttct ggctgaccct gcactttgag ggccgcaact acgaggccag
      841 tgtagacagt ctcaccttca gtgtcgtgac cggcccagcc cctagccagg aggccgggac
      901 caaggcccgt tttccactaa gagatgctgt ggaggagggt gactggacag ccaccgtggt
      961 ggaccagcaa gactgcaccc tctcgctgca gctcaccacc ccggccaacg cccccatcgg
     1021 cctgtatcgc ctcagcctgg aggcctccac tggctaccag ggatccagct ttgtgctggg
     1081 ccacttcatt ttgctcttca acgcctggtg cccagcggat gctgtgtacc tggactcgga
     1141 agaggagcgg caggagtatg tcctcaccca gcagggcttt atctaccagg gctcggccaa
     1201 gttcatcaag aacatacctt ggaattttgg gcagtttgaa gatgggatcc tagacatctg
     1261 cctgatcctt ctagatgtca accccaagtt cctgaagaac gccggccgtg actgctcccg
     1321 ccgcagcagc cccgtctacg tgggccgggt ggtgagtggc atggtcaact gcaacgatga
     1381 ccagggtgtg ctgctgggac gctgggacaa caactacggg gacggcgtca gccccatgtc
     1441 ctggatcggc agcgtggaca tcctgcggcg ctggaagaac cacggctgcc agcgcgtcaa
     1501 gtatggccag tgctgggtct tcgccgccgt ggcctgcaca gtgctgaggt gcctgggcat
     1561 ccctacccgc gtcgtgacca actacaactc ggcccatgac cagaacagca accttctcat
     1621 cgagtacttc cgcaatgagt ttggggagat ccagggtgac aagagcgaga tgatctggaa
     1681 cttccactgc tgggtggagt cgtggatgac caggccggac ctgcagccgg ggtacgaggg
     1741 ctggcaggcc ctggacccaa cgccccagga gaagagcgaa gggacgtact gctgtggccc
     1801 agttccagtt cgtgccatca aggagggcga cctgagcacc aagtacgatg cgccctttgt
     1861 ctttgcggag gtcaatgccg acgtggtaga ctggatccag caggacgatg ggtctgtgca
     1921 caaatccatc aaccgttccc tgatcgttgg gctgaagatc agcactaaga gcgtgggccg
     1981 agacgagcgg gaggatatca cccacaccta caaataccca gaggggtcct cagaggagag
     2041 ggaggccttc acaagggcga accacctgaa caaactggcc gagaaggagg agacagggat
     2101 ggccatgcgg atccgtgtgg gccagagcat gaacatgggc agtgactttg acgtctttgc
     2161 ccacatcacc aacaacaccg ctgaggagta cgtctgccgc ctcctgctct gtgcccgcac
     2221 cgtcagctac aatgggatct tggggcccga gtgtggcacc aagtacctgc tcaacctcaa
     2281 cctggagcct ttctctgaga agagcgttcc tctttgcatc ctctatgaga aataccgtga
     2341 ctgccttacg gagtccaacc tcatcaaggt gcgggccctc ctcgtggagc cagttatcaa
     2401 cagctacctg ctggctgaga gggacctcta cctggagaat ccagaaatca agatccggat
     2461 ccttggggag cccaagcaga aacgcaagct ggtggctgag gtgtccctgc agaacccgct
     2521 ccctgtggcc ctggaaggct gcaccttcac tgtggagggg gccggcctga ctgaggagca
     2581 gaagacggtg gagatcccag accccgtgga ggcaggggag gaagttaagg tgagaatgga
     2641 cctgctgccg ctccacatgg gcctccacaa gctggtggtg aacttcgaga gcgacaagct
     2701 gaaggctgtg aagggcttcc ggaatgtcat cattggcccc gccgaattct gcagatatcc
     2761 agcacagtgg cggccgctcg aggactacaa agaccatgac ggtgattata aagatcatga
     2821 catcgactac aaggatgacg atgacaagta gtgagggccc gatatctcga caatcaacct
     2881 ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg
     2941 ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc
     3001 attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt
     3061 gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc
     3121 attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg
     3181 gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact
     3241 gacaattccg tggtgttgtc ggggaagctg acgtcctttc catggctgct cgcctgtgtt
     3301 gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg
     3361 gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc
     3421 cctcagacga gtcggatctc cctttgggcc gcctccccgc ctggaaacgg gggaggctaa
     3481 ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca ataaaaagac
     3541 agaataaaac gcacgggtgt tgggtcgttt gttcataaac gcggggttcg gtcccagggc
     3601 tggcactctg tcgatacccc accgagaccc cattggggcc aatacgcccg cgtttcttcc
     3661 ttttccccac cccacccccc aagttcgggt gaaggcccag ggctcgcagc caacgtcggg
     3721 gcggcaggcc ctgccatagc agatctgcgc agctggggct ctagggggta tccccacgcg
     3781 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     3841 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     3901 gccggctttc cccgtcaagc tctaaatcgg ggcatccctt tagggttccg atttagtgct
     3961 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     4021 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     4081 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     4141 attttgggga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     4201 aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     4261 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     4321 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     4381 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     4441 atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat
     4501 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag
     4561 cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt cgcatgattg
     4621 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg
     4681 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg
     4741 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg
     4801 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg
     4861 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc
     4921 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc
     4981 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc
     5041 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc
     5101 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg
     5161 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct
     5221 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt
     5281 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc
     5341 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt
     5401 tcttctgagc gggactctgg ggttcgcgaa atgaccgacc aagcgacgcc caacctgcca
     5461 tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc
     5521 cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac
     5581 cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     5641 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     5701 tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag
     5761 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     5821 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     5881 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     5941 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     6001 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     6061 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     6121 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     6181 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     6241 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     6301 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca atgctcacgc
     6361 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     6421 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     6481 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     6541 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca
     6601 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     6661 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     6721 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     6781 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     6841 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     6901 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     6961 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     7021 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     7081 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     7141 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     7201 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     7261 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     7321 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     7381 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     7441 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     7501 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     7561 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     7621 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     7681 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     7741 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     7801 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     7861 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt cgacggatcg
     7921 ggagatctcc cgatccccta tggtcgactc tcagtacaat ctgctctgat gccgcatagt
     7981 taagccagta tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa
     8041 tttaagctac aacaaggcaa ggcttgaccg acaattgcat gaagaatctg cttagg
//
