LOCUS       V046477                10155 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V046477
VERSION     V046477
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..4835
                     /label="Tmem94(NM_028014)"
                     /note="Tmem94(NM_028014)"
                     /gene="Tmem94"
     CDS             4842..4907
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    4931..5519
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    5585..5633
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      5835..6263
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        6277..6606
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             6673..7467
                     /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    7643..7764
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(7813..7829)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7837..7853
                     /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(7861..7891)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7906..7927
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(8215..8803)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8974..9834)
                     /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(9835..9939)
                     /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 ccaccatgga cctgagggag aagcacctgg
      781 gagagccccc cttggccctg ggcctgtcta cccggaaggc cctcagtgtc ctgaaggagc
      841 agctggaggc tgtattggag aaacacctca aagaacggaa gaaaagtctt acatggaagg
      901 aggcatggag aagcagcttc ctgcacctca gtaaccgctg ctcctgtttc cactggcccg
      961 gggcctccct gatgctgctg gcggtgctgc tgctgttgtg ttgctgtggg ggccagccag
     1021 ctgggagcca gggcgtggag ctggtgaacg cctctgctct gttcctgttg ctgctcctca
     1081 acctcgtcct catcgggcgg caagaccggc tgaagcgccg ggaggtggag cgcaggctcc
     1141 gggggatcat cgaccaaatc caagatgctc tcagggatgg aaaagagatc aagtggccga
     1201 actccatgta tccagacctc catatgccct ttgcaccttc ctggtccctg cactgggctt
     1261 acagagatgg acacctggtg aacctgccag ttagcctgtt ggtagaagga gacatcatag
     1321 ccctgaggcc tggccaggag tcctttgcct ccctgagggg catcaaggac gatgagcaca
     1381 ttgtcttgga gccgggagac ctgtttcccc ccttctctcc accaccatcg ccccggggag
     1441 aagtaaagag agggccacag aacccccagc aacaccggct cttccgagtc ctggagacgc
     1501 ctgtgattga caacatcaga tggtgcctgg acacagccct gtcccgcccc gtcacggctc
     1561 tggacaatga aaggttcaca gtacagtcgg tgatgcttca ctatgccgtg ccggtggtcc
     1621 tggctggctt tctcatcacc aatgccctgc gcttcatgtt caaggcacct ggggtcacgt
     1681 cctggcagta cacccttctc cagctccagg tgaacggcat gttgcccatc ctccccctgc
     1741 tcttcccggt cctctgggtg ctggcgaccg cctgtggaga agctcgtgtc ctggcccaga
     1801 tgagcaaggc ttcccccagc tccctgctgg ccaagttctc agaggacact ctcagcagct
     1861 acactgaagc cgtctcctct caggagatgc tgcgctgcat ttggggtcac tttctgaggg
     1921 tgatccaggg cacatcgcca actctgagcc acagtgccag tctgctgcac agcttgggct
     1981 ccgttacggt actgtgctgt gtagacaaac aggggatcct atcatggcca aaccccagcc
     2041 cagagaccgt gctcttcttc agtgggaagg tggagccccc tcacagcagc cacgaggacc
     2101 tcacagacga cctgtccacc cgttccttct gccatcccga ggtggaggag gagccccatg
     2161 aacacgatgc cctccttgct ggctccctga acaataccct gcacctttcc aacgagcagg
     2221 agcgcagtga ctggcttgcc gacggcccca agccctctga gccctaccct catcataaag
     2281 gacatggccg cagcaaacac ccgtctggct ccaacgtgag tttcagcagg gacactgaag
     2341 gcggagagga agagcccagc aaggctcagc ctgggacaga gggtgatccc tacgaggccg
     2401 aggacttcgt gtgtgactac cacttggaga tgctgagcct gtcccaggac caacagaacc
     2461 cctcgtgcat acagttcgat gattccaact ggcagtcaca cctgacctcc ctcaagccgc
     2521 tgggcctcaa cgtgctgctg aacctgtgta acgccagtgt cactgagcgc ctgtgccgct
     2581 tttccgacca cctgtgcaac atcgccctgc aggagagcca cagcgctgtg ctgcccgtgc
     2641 acgtgccctg gggcctctgc gagctggccc ggctcatagg tttcactcca ggggccaagg
     2701 agctcttcaa gcaggagaac cacctggcgc tctaccgcct ccccagtgca gagaccctga
     2761 aggagacgtc attggggagg ccctcctgtg tcaccaagcg gcgccccccg ctcagccaca
     2821 tgatcagcct cttcatcaaa gacactgcca ccagcacaga gcagatgctg tcccatggca
     2881 gtgccgacgt ggtcgtggag gcctgcacag acttctggga tggcgctgac atctaccctc
     2941 tgtcgggttc agacagaaag aaagtgctgg acttctacca gcgagcctgc ctatccggct
     3001 attgctctgc ctttgcctac aagcccatga actgcacgct gtcctctcag ctcaacggca
     3061 agtgcatcga gctggtgcag gtccccggcc agaacagcat attcaccatg tgcgagctgc
     3121 ccagcaccat ccccatcaag ccaaacaacc gccgcagcag ctggagctcc gatgaaggga
     3181 tcggggaggt gctggagaaa gaagactgca tgcaggccct gagcggtcag atcttcatgg
     3241 gcatggtgtc ctcccagtac caggcccggc tggacatcgt gcgcctcatc gatgggctgg
     3301 tcaacgcctg catccgcttt gtgtacttct ctttggaaga tgagctcagg agcaaggtgt
     3361 ttgcagagaa gatgggcctg gagacaggct ggaactgcca catctccctc acgcccaacg
     3421 gtgacatgcc tggctctgag atccctccct ccagccccag ccatgctggc tctctccatg
     3481 acgacctgaa tcaggtgtcc cgagatgatg cagaagggct cctcctctta gaggaggagg
     3541 gtcactcaga cctcatcagc ttccagccca ccgacagcga tatccccagc ttcctggaag
     3601 actgcaaccg ggccaagctg ccccggggca tccaccaggt gcggccccac ctacagaaca
     3661 tcgacaatgt accgttgctg gtgcccctct tcaccgactg cacccctgac accatgtgtg
     3721 agatgataaa gatcatgcag gagtatgggg aggtgacttg ctgcctgggg agctcggcca
     3781 acctgcgcaa cagctgcctc ttcctccaga gtgatgtcag cattgccctg gatcccttat
     3841 acccatcccg ctgctcctgg gagacctttg gctatgccac cagcaccacc atggcccagg
     3901 cctcggatgg cctttctccc ctgcagctat cagggcagct caacagcctg ccttgctccc
     3961 tgaccttccg acaagaagaa agcatcagca tcatccggct cattgagcag gctcggcacg
     4021 ctacctacgg catccgaaag tgcttcctct tcctgttgca gtgccagctg accctcgtgg
     4081 tcattcagtt cctgtcttgt ctggtgcagc tgccaccgct cctgagtacc acagacattt
     4141 tatggctatc ctgcttttgt taccctctgc tcagcatctc tctattggga aaaccaccac
     4201 atagctccat catgtctatg gcaacgggca aaaaccttca gtccattcct aagaagaccc
     4261 agcactactt cctgctctgc ttcctgctca agttcagcct caccatcagc tcctgcctcg
     4321 tctgcttcgg cttcacgctg cagagctttt gtgacagcgc ccgggcccgc aacctcacca
     4381 actgctcctc agtcatgctt tgcagcaatg atgacagagc gccggcctgg ttcgaggatt
     4441 ttgccaatgg gttgctatcc gctcagaagc tcacagccgc cctgattgtt ttgcacactg
     4501 tctttatctc catcactcat gtacaccgca ccaagcctct gtggagaaag agccctttga
     4561 caaacctctg gtgggccgtg actgtgcctg tggtgctgct cggtcaggtg gtccagacag
     4621 tggtggacct gcagctgtgg acacacaggg actcacgtgt ccactttggc ctagaagatg
     4681 tgcccctgct cacatggctc ctgggctgcc tgtccctggt cctagtggta gtcaccaacg
     4741 agattgtaaa gttacacgag attcgggtcc gtgtccgcta ccagaagcgc cagaagctgc
     4801 agtttgagac caagctgggc atgaactcgc ccttcggatc cgactacaaa gaccatgacg
     4861 gtgattataa agatcatgac atcgactaca aggatgacga tgacaagtag tgagggcccg
     4921 atatctcgac aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa
     4981 ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat
     5041 tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta
     5101 tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc
     5161 aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt
     5221 ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg
     5281 ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaagctga cgtcctttcc
     5341 atggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc
     5401 ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct
     5461 tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgcc
     5521 tggaaacggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg aacccgcgct
     5581 atgacggcaa taaaaagaca gaataaaacg cacgggtgtt gggtcgtttg ttcataaacg
     5641 cggggttcgg tcccagggct ggcactctgt cgatacccca ccgagacccc attggggcca
     5701 atacgcccgc gtttcttcct tttccccacc ccacccccca agttcgggtg aaggcccagg
     5761 gctcgcagcc aacgtcgggg cggcaggccc tgccatagca gatctgcgca gctggggctc
     5821 tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     5881 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
     5941 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg gcatcccttt
     6001 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg
     6061 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     6121 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta
     6181 ttcttttgat ttataaggga ttttggggat ttcggcctat tggttaaaaa atgagctgat
     6241 ttaacaaaaa tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag
     6301 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     6361 aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     6421 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     6481 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     6541 gcctctgcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
     6601 tgcaaaaagc tcccgggagc ttgtatatcc attttcggat ctgatcaaga gacaggatga
     6661 ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
     6721 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
     6781 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
     6841 ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
     6901 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
     6961 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
     7021 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
     7081 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
     7141 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
     7201 cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc
     7261 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
     7321 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
     7381 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
     7441 tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgcgaaa tgaccgacca
     7501 agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt
     7561 gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat
     7621 gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag
     7681 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     7741 gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt
     7801 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     7861 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     7921 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     7981 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     8041 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     8101 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     8161 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     8221 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     8281 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     8341 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     8401 gctttctcaa tgctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     8461 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     8521 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     8581 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     8641 cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     8701 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     8761 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     8821 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     8881 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     8941 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     9001 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     9061 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     9121 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     9181 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     9241 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     9301 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     9361 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     9421 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     9481 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     9541 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     9601 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     9661 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     9721 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     9781 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     9841 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     9901 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     9961 acctgacgtc gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc
    10021 tgctctgatg ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct
    10081 gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg
    10141 aagaatctgc ttagg
//
