LOCUS       V021850                 8156 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V021850
VERSION     V021850
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..2803
                     /label="TMEM260(NM_017799)"
                     /note="TMEM260(NM_017799)"
                     /gene="TMEM260"
     CDS             2843..2908
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2932..3520
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3586..3634
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3836..4264
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4278..4607
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4674..5468
                     /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    5644..5765
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5814..5830)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5838..5854
                     /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(5862..5892)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5907..5928
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6216..6804)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6975..7835)
                     /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(7836..7940)
                     /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 ccatgagtcc ccatggcgac ggcaggggcc aggcccaggg
      721 gcgggcagtc cgagtggggc tgcggcgctc cgggggcatc cgcggcggcg tggcggtgtt
      781 cgccgccgtg gccgcagtgt tcaccttcac cctgccccct tcggtaccgg ggggagactc
      841 cggggaactg atcacagccg cacatgagct tggagttgcc catcctcctg gctatccttt
      901 gttcacgctg gtggctaaac tggcaattac actgtttcct tttggttcaa ttgcctaccg
      961 cgtcaatctt ctctgtggct tatttggagc agtagctgca tcattacttt ttttcaccgt
     1021 tttcaggctt tctggctcat ctgctggagg aatccttgct gcgggggtgt tttcattttc
     1081 tcgtctaaca tggcagtggt ccattgcagc agaggttttt agcttaaaca atctctttgt
     1141 ggggctgctt atggctctta ctgtacattt tgaggaagca gcaactgcta aggagagatc
     1201 aaaggtagct aaaattggtg ctttctgctg tggccttagt ttatgtaacc agcacacaat
     1261 aatactctat gttttgtgca taataccttg gattctcttt caacttttaa aaaagaagga
     1321 actctccctg ggctctttgt tgaagttgag cctgtacttc tctgctggtt tgctgcccta
     1381 tgtccacctt cccatctcat cttaccttaa tcacgcccgg tggacctggg gagaccagac
     1441 aacactgcaa ggatttttga cacattttct cagggaagaa tatggaacct tcagcctggc
     1501 caaatctgaa ataggatcca gtatgtctga aatactgctt tctcaagtaa caaatatgag
     1561 gaccgaactc tcattcaaca tccaagccct tgcagtttgt gcaaatatat gtttagcaac
     1621 aaaggacaga cagaatccat cattagtatg gctttttact ggaatgtttt gcatttattc
     1681 attgttcttt gcttggagag caaatttaga tatttcaaaa ccacttttca tgggtgtggt
     1741 ggaacgattc tggatgcaga gcaatgcagt agtggccgtc ctcgctggca ttggtttggc
     1801 tgcagttgtg tctgagacta accgagtgct gaatagcaat gggcttcagt gtctggaatg
     1861 gctttctgca actctttttg tagtttacca aatatattct aattacagtg tttgtgacca
     1921 aaggaccaac tatgtgattg ataagttcgc aaagaacctt ctcacctcta tgcctcatga
     1981 tgcaattatc ttactcagag gagatttgcc aggaaattct ctccgttaca tgcattactg
     2041 tgaggggttg aggcctgaca tttcattagt ggatcaggaa atgatgactt acgagtggta
     2101 tttacccaag atggcaaagc acttgccagg tgtcaacttt cctgggaacc ggtggaatcc
     2161 tgtggaagga atattaccta gtggaatggt cacatttaat ctttatcatt ttcttgaagt
     2221 aaataaacaa aaagaaacat ttgtttgcat aggaattcat gaaggcgacc caacctggaa
     2281 aaagaactat tcactttggc catgggggtc ttgtgacaaa ttagttcctt tggagattgt
     2341 attcaaccct gaggaatgga ttaaacttac aaaaagtatc tataactgga ccgaagaata
     2401 tggaaggttt gatccatctt cttgggaatc tgtggccaat gaagaaatgt ggcaagcgag
     2461 gatgaaaaca ccgttcttca tctttaacct ggcagaaact gctcacatgc cttcaaaagt
     2521 gaaagctcaa ctctacgctc aagcatatga cctttataag gagattgtct atttacaaaa
     2581 ggagcaccca gtgaattggc acaagaacta tgccatcgcc tgtgagcgga tgctgcgtct
     2641 tcaggcaaga gatgcagatc ctgaagtgct gttatcggaa accatcagac atttccgtct
     2701 gtactctcag aaagcaccga atgacccaca gcaagctgat attttaggtg ctctaaagca
     2761 cctaagaaaa gaactgcaaa gtctgagaaa taggaaaaat gtcgaattct gcagatatcc
     2821 agcacagtgg cggccgctcg aggactacaa agaccatgac ggtgattata aagatcatga
     2881 catcgactac aaggatgacg atgacaagta gtgagggccc gatatctcga caatcaacct
     2941 ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg
     3001 ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc
     3061 attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt
     3121 gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc
     3181 attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg
     3241 gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact
     3301 gacaattccg tggtgttgtc ggggaagctg acgtcctttc catggctgct cgcctgtgtt
     3361 gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg
     3421 gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc
     3481 cctcagacga gtcggatctc cctttgggcc gcctccccgc ctggaaacgg gggaggctaa
     3541 ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca ataaaaagac
     3601 agaataaaac gcacgggtgt tgggtcgttt gttcataaac gcggggttcg gtcccagggc
     3661 tggcactctg tcgatacccc accgagaccc cattggggcc aatacgcccg cgtttcttcc
     3721 ttttccccac cccacccccc aagttcgggt gaaggcccag ggctcgcagc caacgtcggg
     3781 gcggcaggcc ctgccatagc agatctgcgc agctggggct ctagggggta tccccacgcg
     3841 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     3901 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     3961 gccggctttc cccgtcaagc tctaaatcgg ggcatccctt tagggttccg atttagtgct
     4021 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     4081 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     4141 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     4201 attttgggga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     4261 aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     4321 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     4381 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     4441 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     4501 atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat
     4561 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag
     4621 cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt cgcatgattg
     4681 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg
     4741 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg
     4801 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg
     4861 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg
     4921 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc
     4981 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc
     5041 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc
     5101 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc
     5161 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg
     5221 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct
     5281 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt
     5341 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc
     5401 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt
     5461 tcttctgagc gggactctgg ggttcgcgaa atgaccgacc aagcgacgcc caacctgcca
     5521 tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc
     5581 cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac
     5641 cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     5701 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     5761 tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag
     5821 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     5881 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     5941 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     6001 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     6061 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     6121 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     6181 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     6241 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     6301 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     6361 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca atgctcacgc
     6421 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     6481 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     6541 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     6601 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca
     6661 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     6721 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     6781 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     6841 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     6901 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     6961 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     7021 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     7081 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     7141 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     7201 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     7261 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     7321 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     7381 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     7441 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     7501 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     7561 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     7621 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     7681 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     7741 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     7801 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     7861 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     7921 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt cgacggatcg
     7981 ggagatctcc cgatccccta tggtcgactc tcagtacaat ctgctctgat gccgcatagt
     8041 taagccagta tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa
     8101 tttaagctac aacaaggcaa ggcttgaccg acaattgcat gaagaatctg cttagg
//
