LOCUS       V027733                 7833 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V027733
VERSION     V027733
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..2513
                     /label="Ezr(NM_009510)"
                     /note="Ezr(NM_009510)"
                     /gene="Ezr"
     CDS             2520..2585
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2609..3197
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3263..3311
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3513..3941
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3955..4284
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4351..5145
                     /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    5321..5442
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5491..5507)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5515..5531
                     /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(5539..5569)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5584..5605
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5893..6481)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6652..7512)
                     /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(7513..7617)
                     /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 ccaccatgcc caagccaatc aacgtccggg
      781 tgaccaccat ggatgccgag ctggagtttg ccatccagcc aaacaccacc gggaagcagc
      841 tctttgatca ggtggtaaag acgattggcc tccgggaagt gtggtacttc ggcctccagt
      901 atgtagacaa taaaggattt cctacctggc tgaaacttga taaaaaggtc tccgcacagg
      961 aggtccgaaa ggagaaccct gtccagttta aattccgggc caagttctac cccgaagacg
     1021 tggccgagga actcatccag gacatcacac agaagctctt cttcctgcaa gtcaaggacg
     1081 ggatcctcag cgacgagatc tactgccccc cagagacagc cgtgctcctg ggctcctatg
     1141 ccgttcaggc caagttcgga gattataaca aggaaatgca caagtctggg tacctcagct
     1201 cggagcggct gatcccccag agagtcatgg accaacacaa gctcagcagg gaccagtggg
     1261 aggaccggat ccaggtgtgg cacgcggaac accgagggat gctcaaggac agtgctatgc
     1321 tagaatacct gaagattgcc caggacctgg aaatgtatgg gatcaactat ttcgagatca
     1381 aaaacaagaa aggaacagac ctttggcttg gagtcgatgc ccttggactt aacatttatg
     1441 agaaagatga caagttgacc ccaaagatcg gcttcccttg gagtgagatc aggaacatct
     1501 ctttcaacga caagaagttt gtcattaagc ccatcgacaa gaaggcacct gactttgtgt
     1561 tctacgcccc gcgcctgaga attaacaagc ggatcctgca gctctgcatg gggaaccatg
     1621 agctgtacat gcgccgcagg aagcccgaca ccatcgaggt gcagcagatg aaggcccagg
     1681 ctcgggagga gaagcaccag aagcagctag agcgacagca gttggaaacc gagaagaaga
     1741 ggcgagagac ggtggagaga gaaaaggagc agatgctccg ggagaaggag gagctgatgc
     1801 ttcggctgca ggactacgag cagaagacca agagggcgga gaaagagctc tccgagcaga
     1861 ttgagaaggc cctccaactg gaggaagaga ggaggcgagc ccaggaggag gctgaacgtc
     1921 tggaggccga ccgcatggcc gccctgcggg ccaaggaaga actcgagaga caggcgcagg
     1981 atcagataaa gagccaggag cagctggctg cagagctggc agagtacacg gccaagatcg
     2041 cactgctgga ggaggcgcgg aggcgcaagg aggacgaggt agaagagtgg cagcaccggg
     2101 ctaaagaagc ccaggacgac ctggtgaaga ccaaagagga gctgcacctg gtgatgacgg
     2161 ccccaccgcc cccaccgccc ccagtgtatg agcctgtgaa ttaccacgtg caggagggac
     2221 tgcaggacga gggagcagag cctatgggct acagtgccga gctctccagt gagggcatcc
     2281 tggatgaccg caacgaggag aagcggatca cagaggcaga gaagaatgag cgcgtgcagc
     2341 ggcagctgct gaccctgagc aatgagttgt cccaggcccg ggatgagaac aagaggaccc
     2401 acaatgacat catccacaac gagaacatgc ggcaaggcag ggacaagtat aagacgctgc
     2461 ggcaaatcag gcagggcaac accaagcaac gcattgacga gttcgaggcc atgggatccg
     2521 actacaaaga ccatgacggt gattataaag atcatgacat cgactacaag gatgacgatg
     2581 acaagtagtg agggcccgat atctcgacaa tcaacctctg gattacaaaa tttgtgaaag
     2641 attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat
     2701 gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc
     2761 ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg
     2821 cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct
     2881 ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct
     2941 tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg
     3001 gaagctgacg tcctttccat ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac
     3061 gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct
     3121 gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct
     3181 ttgggccgcc tccccgcctg gaaacggggg aggctaactg aaacacggaa ggagacaata
     3241 ccggaaggaa cccgcgctat gacggcaata aaaagacaga ataaaacgca cgggtgttgg
     3301 gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg cactctgtcg ataccccacc
     3361 gagaccccat tggggccaat acgcccgcgt ttcttccttt tccccacccc accccccaag
     3421 ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg ccatagcaga
     3481 tctgcgcagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc
     3541 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     3601 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     3661 aaatcggggc atccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     3721 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     3781 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     3841 caaccctatc tcggtctatt cttttgattt ataagggatt ttggggattt cggcctattg
     3901 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt
     3961 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     4021 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     4081 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     4141 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     4201 tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt
     4261 tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct
     4321 gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt
     4381 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc
     4441 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag
     4501 accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg
     4561 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac
     4621 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc
     4681 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc
     4741 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc
     4801 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg
     4861 ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat
     4921 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc
     4981 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa
     5041 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat
     5101 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt
     5161 tcgcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc
     5221 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct
     5281 ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta
     5341 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     5401 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc
     5461 gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     5521 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     5581 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     5641 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     5701 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     5761 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     5821 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     5881 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     5941 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     6001 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     6061 cccttcggga agcgtggcgc tttctcaatg ctcacgctgt aggtatctca gttcggtgta
     6121 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     6181 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     6241 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     6301 gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct
     6361 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     6421 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     6481 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     6541 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     6601 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     6661 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     6721 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     6781 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     6841 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     6901 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     6961 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     7021 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     7081 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     7141 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     7201 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     7261 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     7321 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     7381 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     7441 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     7501 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     7561 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     7621 atttccccga aaagtgccac ctgacgtcga cggatcggga gatctcccga tcccctatgg
     7681 tcgactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatct gctccctgct
     7741 tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc
     7801 ttgaccgaca attgcatgaa gaatctgctt agg
//
