LOCUS       V043148                 7356 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V043148
VERSION     V043148
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..2036
                     /label="PUS1(NM_025215)"
                     /note="PUS1(NM_025215)"
                     /gene="PUS1"
     CDS             2043..2108
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2132..2720
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2786..2834
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3036..3464
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3478..3807
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3874..4668
                     /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    4844..4965
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5014..5030)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5038..5054
                     /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(5062..5092)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5107..5128
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5416..6004)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6175..7035)
                     /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(7036..7140)
                     /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 ccaccatggg cctccagctt cgcgcgctgt
      781 tgggagcctt cggacggtgg accctgcgcc tgggaccgcg tccgtcctgc tcgccgcgca
      841 tggccgggaa cgcggagccg ccgcccgccg gagccgcatg cccccaggac cggaggtcct
      901 gcagcggccg ggccgggggc gaccgcgtct gggaggacgg agaacatccg gcgaagaagc
      961 tcaagagcgg tggcgacgag gagcggcgcg agaagccgcc caagcggaag atcgtgctgc
     1021 tcatggccta ttcgggcaag ggctaccacg gcatgcagag gaatgtcggg tcctcacaat
     1081 tcaaaacaat tgaagatgac ttggtgtccg ccctcgtccg gtcaggctgt attcctgaaa
     1141 atcatggtga ggacatgagg aaaatgtcct tccagcgctg cgcccggaca gacaagggtg
     1201 tgtccgcagc cggccaggtg gtatccctga aggtgtggct gattgacgac attctagaaa
     1261 agatcaacag ccaccttccc tctcacattc ggattctggg actgaagcgg gtcacgggcg
     1321 ggtttaactc caagaacaga tgtgatgcca ggacctattg ctacctgctg cccacgtttg
     1381 cctttgcgca caaggaccgg gacgttcagg atgagaccta ccgcctgagc gccgagacgc
     1441 tgcagcaggt caacaggctc ctggcctgct acaagggcac gcacaacttc cacaatttca
     1501 cctcgcagaa ggggccgcag gatcccagtg cctgccgcta catcctggag atgtactgcg
     1561 aggaaccctt tgtgcgggag ggcctggagt ttgcggtgat cagggtgaag ggccagagct
     1621 tcatgatgca tcagatccgg aagatggtcg gcctggtggt ggccattgtg aagggttatg
     1681 cccctgagag cgtgctggag cgcagctggg gcacagagaa ggtggacgtg cccaaggcgc
     1741 ccggactcgg cctggtcctg gagagggtgc acttcgagaa gtacaaccag cgctttggca
     1801 acgatgggct gcatgagccg ctggactggg cgcaggagga aggaaaggtc gcagccttca
     1861 aggaggagca catctacccc accatcatcg gcaccgagcg ggacgaacgc tccatggccc
     1921 agtggctgag caccttgccc atccacaact tcagtgccac cgctctcacg gcaggtggca
     1981 cgggcgccaa ggtgcccagt cccctggaag gcagtgaagg ggacggagac actgacggat
     2041 ccgactacaa agaccatgac ggtgattata aagatcatga catcgactac aaggatgacg
     2101 atgacaagta gtgagggccc gatatctcga caatcaacct ctggattaca aaatttgtga
     2161 aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt
     2221 aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa
     2281 atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt
     2341 gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct
     2401 cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg
     2461 ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc
     2521 ggggaagctg acgtcctttc catggctgct cgcctgtgtt gccacctgga ttctgcgcgg
     2581 gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct
     2641 gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc
     2701 cctttgggcc gcctccccgc ctggaaacgg gggaggctaa ctgaaacacg gaaggagaca
     2761 ataccggaag gaacccgcgc tatgacggca ataaaaagac agaataaaac gcacgggtgt
     2821 tgggtcgttt gttcataaac gcggggttcg gtcccagggc tggcactctg tcgatacccc
     2881 accgagaccc cattggggcc aatacgcccg cgtttcttcc ttttccccac cccacccccc
     2941 aagttcgggt gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc
     3001 agatctgcgc agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag
     3061 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     3121 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     3181 tctaaatcgg ggcatccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     3241 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     3301 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     3361 actcaaccct atctcggtct attcttttga tttataaggg attttgggga tttcggccta
     3421 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg
     3481 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     3541 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
     3601 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
     3661 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     3721 atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc
     3781 ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga
     3841 tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca
     3901 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc
     3961 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc
     4021 aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg
     4081 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg
     4141 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct
     4201 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct
     4261 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa
     4321 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa
     4381 ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc
     4441 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt
     4501 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct
     4561 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc
     4621 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg
     4681 ggttcgcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac
     4741 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat
     4801 cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc
     4861 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     4921 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc
     4981 gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     5041 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     5101 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     5161 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     5221 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     5281 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     5341 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     5401 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     5461 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     5521 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     5581 tctcccttcg ggaagcgtgg cgctttctca atgctcacgc tgtaggtatc tcagttcggt
     5641 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     5701 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     5761 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     5821 cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct
     5881 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     5941 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     6001 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     6061 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     6121 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     6181 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     6241 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     6301 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     6361 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     6421 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     6481 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     6541 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     6601 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     6661 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     6721 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     6781 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     6841 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     6901 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     6961 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     7021 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     7081 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     7141 cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc cgatccccta
     7201 tggtcgactc tcagtacaat ctgctctgat gccgcatagt taagccagta tctgctccct
     7261 gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa
     7321 ggcttgaccg acaattgcat gaagaatctg cttagg
//
