LOCUS       V021070                 9599 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V021070
VERSION     V021070
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..4279
                     /label="TTLL4(NM_014640)"
                     /note="TTLL4(NM_014640)"
                     /gene="TTLL4"
     CDS             4286..4351
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    4375..4963
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    5029..5077
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      5279..5707
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5721..6050
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             6117..6911
                     /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    7087..7208
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(7257..7273)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7281..7297
                     /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(7305..7335)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7350..7371
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7659..8247)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8418..9278)
                     /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(9279..9383)
                     /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 ccatggcctc agcaggaaca cagcactata gtattggcct
      721 ccgccagaaa aacagcttca agcagagtgg tccctcaggc acagtacctg ccacgccacc
      781 tgagaaaccc tcggagggca gagtctggcc tcaggcccat cagcaagtga agccaatctg
      841 gaagctggaa aagaagcaag tggagacact gtcagcaggg ttgggcccag gcctcttggg
      901 cgtcccaccc cagccagcat atttcttttg ccccagcact ttatgtagct ctgggaccac
      961 ggctgtcatt gcaggccaca gcagttcctg ttacctacac tctctcccgg acttgttcaa
     1021 cagcaccctg ctataccgcc gctccagcta taggcaaaaa ccgtaccagc aactggagtc
     1081 tttctgcttg cgttcgagcc cgtcagaaaa aagccctttt tctctccctc aaaagagcct
     1141 ccctgtcagt ctcactgcca acaaggccac ttcttccatg gtcttctcca tggcccagcc
     1201 catggcctcc tcatccacag aaccatacct ctgcttggca gcggctgggg aaaacccttc
     1261 agggaagagc ctggcctctg ccatctcagg gaagatccca tctccactct cttcctccta
     1321 taagcccatg ctgaataata attccttcat gtggccaaat agcacgccag tgcctttatt
     1381 gcagaccaca cagggcctga agccagtatc gccacccaag atccagcctg tctcctggca
     1441 tcattcaggg ggtactggag actgtgcacc gcagcctgtt gaccataagg tgcccaaaag
     1501 cattggcact gtcccagctg atgccagtgc ccatatcgcc ttgtctaccg ctagctccca
     1561 cgacacatcc accaccagtg ttgcctcttc ctggtataac cggaataact tagccatgag
     1621 ggcagagcca ctttcctgtg ctctggatga cagctctgat tcccaggatc caactaagga
     1681 gattcggttc actgaggccg tgaggaaatt gaccgcaaga ggctttgaga agatgccgag
     1741 gcaaggctgc cagcttgaac agtctagttt cctgaacccc agcttccagt ggaatgtcct
     1801 caacaggagc aggcggtgga aacctcctgc ggtaaatcag cagtttcctc aggaggatgc
     1861 tggatcggtc aggcgggtcc tccctggtgc ctcagatacc ttggggttgg acaatacagt
     1921 cttctgtacc aagcgtatca gcattcacct ccttgcctca catgccagtg ggctcaatca
     1981 caaccctgcc tgtgaatctg taattgactc ctcagcattt ggagaaggca aagctccagg
     2041 tccccctttt cctcaaactc ttggcatagc caacgtggcc acccgcctct cttccatcca
     2101 gctgggccag tctgagaagg agagacctga ggaggccagg gagctggact catctgatag
     2161 ggatattagt tcagctactg acctccagcc agatcaggct gagactgaag atacagaaga
     2221 agaactagta gatggtttgg aagactgttg tagccgtgat gagaatgaag aggaggaggg
     2281 agactcagag tgctcctcat taagtgctgt ctcccccagc gaatcggtgg ccatgatctc
     2341 tagaagctgt atggaaattc tgaccaaacc cctttccaat catgagaaag ttgtccgacc
     2401 agccctcatc tacagtctct ttcccaacgt tccccctacc atctattttg gcactcggga
     2461 tgagagagtg gagaaacttc cctgggaaca gaggaagttg ctccgatgga agatgagcac
     2521 agtgaccccc aacattgtca agcagaccat tggacggtcc cacttcaaaa tcagcaaaag
     2581 aaacgatgac tggctgggct gctggggtca ccacatgaag tctcctagtt tccgatccat
     2641 tcgagagcat cagaagctaa accatttccc aggctcattc cagattggga ggaaggaccg
     2701 gctatggcgg aacctgtcac gtatgcagag ccgctttggc aagaaggagt tcagtttctt
     2761 cccccagtcc tttatcctgc cccaggacgc caagctcctg cgcaaagcgt gggagagcag
     2821 cagccgccaa aagtggattg tgaagccacc agcatcagct cgaggcattg gcatccaggt
     2881 tattcacaag tggagtcagc tccccaagcg aaggcccctc ctggtacaga ggtatctaca
     2941 caaaccctac ctcatcagcg gcagcaagtt tgacctgcgg atctatgttt atgtcacttc
     3001 ctacgatcct ctgcggattt acctcttttc agatggactg gtccgctttg ccagttgcaa
     3061 gtattcgcct tccatgaaga gccttggcaa taagttcatg cacctgacca actacagtgt
     3121 caataaaaag aatgccgagt accaggccaa tgcagatgaa atggcttgcc agggccacaa
     3181 atgggcactg aaggctttgt ggaactacct gagccagaag ggagtcaata gcgacgccat
     3241 ctgggagaag ataaaggatg ttgttgtcaa aactatcatc tcgtcagagc cctatgtgac
     3301 cagcctgctc aagatgtatg tgcgacggcc ctatagctgc catgaactct ttggttttga
     3361 catcatgcta gacgaaaacc tcaagccctg ggtcctggaa gtcaacattt ccccaagcct
     3421 ccactccagc tctccactgg atatcagcat caaaggccag atgattcgtg accttctgaa
     3481 tctggcaggt tttgtcctgc ccaatgcaga ggatatcatt tccagcccca gcagctgcag
     3541 cagctccacc accagcctgc ccacctcccc tggggacaaa tgtcgaatgg ctccagagca
     3601 tgtcactgca cagaagatga agaaagccta ttatctgacc cagaaaattc ctgatcagga
     3661 cttctatgca tctgtgctgg atgtcctgac accagatgat gttcggattc tggttgagat
     3721 ggaagatgag ttttctcgcc gtggtcagtt tgaacgaatt tttccttctc atatctcctc
     3781 tcgctatctc cgcttttttg agcagccacg atatttcaac attctcacca cccaatggga
     3841 acagaaatac catggcaaca agcttaaagg agtagatctg ctccggagtt ggtgctacaa
     3901 agggttccac atgggagttg tctctgattc tgctccagtg tggtctctcc cgacatcact
     3961 tctgactatc tcaaaggatg acgtgatact caatgccttc agcaaatcag agactagcaa
     4021 gctgggaaaa caaagctcct gtgaggttag cctactactc tctgaagacg ggaccacgcc
     4081 caaatccaag aagactcaag ctggcctttc cccttatccc cagaaaccca gttcctcaaa
     4141 ggacagtgag gacaccagca aagagcccag cctttctacc cagacgttac ctgtgatcaa
     4201 gtgctctggg cagacttcaa gactttctgc ttcctccact ttccagtcaa tcagtgactc
     4261 cctcctggct gtgagcccac tcgaggacta caaagaccat gacggtgatt ataaagatca
     4321 tgacatcgac tacaaggatg acgatgacaa gtagtgaggg cccgatatct cgacaatcaa
     4381 cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt
     4441 acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct
     4501 ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc
     4561 gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg
     4621 ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc
     4681 acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc
     4741 actgacaatt ccgtggtgtt gtcggggaag ctgacgtcct ttccatggct gctcgcctgt
     4801 gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca
     4861 gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt
     4921 cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcctggaaa cgggggaggc
     4981 taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa
     5041 gacagaataa aacgcacggg tgttgggtcg tttgttcata aacgcggggt tcggtcccag
     5101 ggctggcact ctgtcgatac cccaccgaga ccccattggg gccaatacgc ccgcgtttct
     5161 tccttttccc caccccaccc cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc
     5221 ggggcggcag gccctgccat agcagatctg cgcagctggg gctctagggg gtatccccac
     5281 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     5341 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     5401 ttcgccggct ttccccgtca agctctaaat cggggcatcc ctttagggtt ccgatttagt
     5461 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     5521 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     5581 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     5641 gggattttgg ggatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     5701 gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     5761 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     5821 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     5881 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     5941 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc
     6001 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg
     6061 gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg tttcgcatga
     6121 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
     6181 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
     6241 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg
     6301 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
     6361 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
     6421 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
     6481 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
     6541 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
     6601 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg
     6661 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
     6721 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
     6781 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
     6841 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
     6901 agttcttctg agcgggactc tggggttcgc gaaatgaccg accaagcgac gcccaacctg
     6961 ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt
     7021 ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc
     7081 caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     7141 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     7201 gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca
     7261 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     7321 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     7381 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     7441 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     7501 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     7561 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     7621 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     7681 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     7741 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     7801 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcaatgctca
     7861 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     7921 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     7981 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     8041 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg
     8101 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     8161 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     8221 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     8281 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     8341 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     8401 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     8461 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     8521 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     8581 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     8641 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     8701 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     8761 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     8821 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     8881 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     8941 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     9001 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     9061 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     9121 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     9181 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     9241 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     9301 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     9361 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacgga
     9421 tcgggagatc tcccgatccc ctatggtcga ctctcagtac aatctgctct gatgccgcat
     9481 agttaagcca gtatctgctc cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca
     9541 aaatttaagc tacaacaagg caaggcttga ccgacaattg catgaagaat ctgcttagg
//
