LOCUS       pEnCMV-TF(human)-MBP    9308 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V033852
VERSION     V033852
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             775..2868
                     /label="TF(NM_001063)"
                     /note="TF(NM_001063)"
                     /gene="TF"
     CDS             2914..4014
                     /codon_start=1
                     /gene="malE (mutated)"
                     /note="This version of the gene does not encode a signal
                     sequence, so MBP will remain in the cytosol."
                     /product="maltose binding protein from E. coli"
                     /transl_table=1
                     /translation="MKIEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLE
                     EKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKL
                     IAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIA
                     ADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGE
                     TAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFL
                     ENYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAF
                     WYAVRTAVINAASGRQTVDEALKDAQT"
                     /label="MBP"
     misc_feature    4084..4672
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4738..4786
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4988..5416
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5430..5759
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5826..6620
                     /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    6796..6917
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6966..6982)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6990..7006
                     /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(7014..7044)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7059..7080
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7368..7956)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8127..8987)
                     /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(8988..9092)
                     /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 gctcgtttag tgaaccgtca gatcgcctgg
      661 agacgccatc cacgctgttt tgacctccat agaagacacc gggaccgatc cagcctccgg
      721 actctagagc tagcgtttaa acttaagctt ggtaccgagc tcggatccgc caccatgagg
      781 ctcgccgtgg gagccctgct ggtctgcgcc gtcctggggc tgtgtctggc tgtccctgat
      841 aaaactgtga gatggtgtgc agtgtcggag catgaggcca ctaagtgcca gagtttccgc
      901 gaccatatga aaagcgtcat tccatccgat ggtcccagtg ttgcttgtgt gaagaaagcc
      961 tcctaccttg attgcatcag ggccattgcg gcaaacgaag cggatgctgt gacactggat
     1021 gcaggtttgg tgtatgatgc ttacctggct cccaataacc tgaagcctgt ggtggcagag
     1081 ttctatgggt caaaagagga tccacagact ttctattatg ctgttgctgt ggtgaagaag
     1141 gatagtggct tccagatgaa ccagcttcga ggcaagaagt cctgccacac gggtctaggc
     1201 aggtccgctg ggtggaacat ccccataggc ttactttact gtgacttacc tgagccacgt
     1261 aaacctcttg agaaagcagt ggccaatttc ttctcgggca gctgtgcccc ttgtgcggat
     1321 gggacggact tcccccagct gtgtcaactg tgtccagggt gtggctgctc cacccttaac
     1381 caatacttcg gctactcggg agccttcaag tgtctgaagg atggtgctgg ggatgtggcc
     1441 tttgtcaagc actcgactat atttgagaac ttggcaaaca aggctgacag ggaccagtat
     1501 gagctgcttt gcctggacaa cacccggaag ccggtagatg aatacaagga ctgccacttg
     1561 gcccaggtcc cttctcatac cgtcgtggcc cgaagtatgg gcggcaagga ggacttgatc
     1621 tgggagcttc tcaaccaggc ccaggaacat tttggcaaag acaaatcaaa agaattccaa
     1681 ctattcagct ctcctcatgg gaaggacctg ctgtttaagg actctgccca cgggttttta
     1741 aaagtccccc ccaggatgga tgccaagatg tacctgggct atgagtatgt cactgccatc
     1801 cggaatctac gggaaggcac atgcccagaa gccccaacag atgaatgcaa gcctgtgaag
     1861 tggtgtgcgc tgagccacca cgagaggctc aagtgtgatg agtggagtgt taacagtgta
     1921 gggaaaatag agtgtgtatc agcagagacc accgaagact gcatcgccaa gatcatgaat
     1981 ggagaagctg atgccatgag cttggatgga gggtttgtct acatagcggg caagtgtggt
     2041 ctggtgcctg tcttggcaga aaactacaat aagagcgata attgtgagga tacaccagag
     2101 gcagggtatt ttgctatagc agtggtgaag aaatcagctt ctgacctcac ctgggacaat
     2161 ctgaaaggca agaagtcctg ccatacggca gttggcagaa ccgctggctg gaacatcccc
     2221 atgggcctgc tctacaataa gatcaaccac tgcagatttg atgaattttt cagtgaaggt
     2281 tgtgcccctg ggtctaagaa agactccagt ctctgtaagc tgtgtatggg ctcaggccta
     2341 aacctgtgtg aacccaacaa caaagaggga tactacggct acacaggcgc tttcaggtgt
     2401 ctggttgaga agggagatgt ggcctttgtg aaacaccaga ctgtcccaca gaacactggg
     2461 ggaaaaaacc ctgatccatg ggctaagaat ctgaatgaaa aagactatga gttgctgtgc
     2521 cttgatggta ccaggaaacc tgtggaggag tatgcgaact gccacctggc cagagccccg
     2581 aatcacgctg tggtcacacg gaaagataag gaagcttgcg tccacaagat attacgtcaa
     2641 cagcagcacc tatttggaag caacgtaact gactgctcgg gcaacttttg tttgttccgg
     2701 tcggaaacca aggaccttct gttcagagat gacacagtat gtttggccaa acttcatgac
     2761 agaaacacat atgaaaaata cttaggagaa gaatatgtca aggctgttgg taacctgaga
     2821 aaatgctcca cctcatcact cctggaagcc tgcactttcc gtagaccttg tacaggccgg
     2881 cctggcggtg gaggcagtgg tggaggcggt agcatgaaaa tcgaagaagg taaactggta
     2941 atctggatta acggcgataa aggctataac ggtctcgctg aagtcggtaa gaaattcgag
     3001 aaagataccg gaattaaagt caccgttgag catccggata aactggaaga gaaattccca
     3061 caggttgcgg caactggcga tggccctgac attatcttct gggcacacga ccgctttggt
     3121 ggctacgctc aatctggcct gttggctgaa atcaccccgg acaaagcgtt ccaggacaag
     3181 ctgtatccgt ttacctggga tgccgtacgt tacaacggca agctgattgc ttacccgatc
     3241 gctgttgaag cgttatcgct gatttataac aaagatctgc tgccgaaccc gccaaaaacc
     3301 tgggaagaga tcccggcgct ggataaagaa ctgaaagcga aaggtaagag cgcgctgatg
     3361 ttcaacctgc aagaaccgta cttcacctgg ccgctgattg ctgctgacgg gggttatgcg
     3421 ttcaagtatg aaaacggcaa gtacgacatt aaagacgtgg gcgtggataa cgctggcgcg
     3481 aaagcgggtc tgaccttcct ggttgacctg attaaaaaca aacacatgaa tgcagacacc
     3541 gattactcca tcgcagaagc tgcctttaat aaaggcgaaa cagcgatgac catcaacggc
     3601 ccgtgggcat ggtccaacat cgacaccagc aaagtgaatt atggtgtaac ggtactgccg
     3661 accttcaagg gtcaaccatc caaaccgttc gttggcgtgc tgagcgcagg tattaacgcc
     3721 gccagtccga acaaagagct ggcaaaagag ttcctcgaaa actatctgct gactgatgaa
     3781 ggtctggaag cggttaataa agacaaaccg ctgggtgccg tagcgctgaa gtcttacgag
     3841 gaagagttgg cgaaagatcc acgtattgcc gccactatgg aaaacgccca gaaaggtgaa
     3901 atcatgccga acatcccgca gatgtccgct ttctggtatg ccgtgcgtac tgcggtgatc
     3961 aacgccgcca gcggtcgtca gactgtcgat gaagccctga aagacgcgca gacttgactc
     4021 gagtctagag ggccccttaa gggttcgatc cctaccggtt agtaatgagt ttgatatctc
     4081 gacaatcaac ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt
     4141 gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc
     4201 cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag
     4261 ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc
     4321 actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc
     4381 cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg
     4441 ctgttgggca ctgacaattc cgtggtgttg tcggggaagc tgacgtcctt tccatggctg
     4501 ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc
     4561 ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt
     4621 cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc gcctggaaac
     4681 gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg
     4741 caataaaaag acagaataaa acgcacgggt gttgggtcgt ttgttcataa acgcggggtt
     4801 cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg ccaatacgcc
     4861 cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc agggctcgca
     4921 gccaacgtcg gggcggcagg ccctgccata gcagatctgc gcagctgggg ctctaggggg
     4981 tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     5041 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     5101 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc ggggcatccc tttagggttc
     5161 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
     5221 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     5281 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
     5341 gatttataag ggattttggg gatttcggcc tattggttaa aaaatgagct gatttaacaa
     5401 aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
     5461 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
     5521 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     5581 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     5641 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg
     5701 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
     5761 agctcccggg agcttgtata tccattttcg gatctgatca agagacagga tgaggatcgt
     5821 ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc
     5881 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc
     5941 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg
     6001 aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag
     6061 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg
     6121 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg
     6181 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac
     6241 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg
     6301 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc
     6361 ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg
     6421 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc
     6481 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc
     6541 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc
     6601 ttcttgacga gttcttctga gcgggactct ggggttcgcg aaatgaccga ccaagcgacg
     6661 cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc
     6721 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag
     6781 ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     6841 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     6901 ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa
     6961 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     7021 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     7081 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     7141 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     7201 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     7261 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     7321 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     7381 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     7441 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     7501 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     7561 caatgctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     7621 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     7681 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     7741 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     7801 actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     7861 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     7921 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     7981 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     8041 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     8101 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     8161 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     8221 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     8281 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     8341 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     8401 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     8461 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     8521 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     8581 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     8641 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     8701 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     8761 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     8821 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     8881 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     8941 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     9001 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     9061 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     9121 gtcgacggat cgggagatct cccgatcccc tatggtcgac tctcagtaca atctgctctg
     9181 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
     9241 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
     9301 tgcttagg
//
