LOCUS       V045844                 7278 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V045844
VERSION     V045844
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..1958
                     /label="PHLDA1(NM_007350)"
                     /note="PHLDA1(NM_007350)"
                     /gene="PHLDA1"
     CDS             1965..2030
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2054..2642
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2708..2756
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      2958..3386
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3400..3729
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3796..4590
                     /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    4766..4887
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4936..4952)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4960..4976
                     /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(4984..5014)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5029..5050
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5338..5926)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6097..6957)
                     /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(6958..7062)
                     /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 ccaccatgag gcgtgcgccg gctgccgagc
      781 gcctcttgga gctgggcttt cccccgcggt gcgggcgcca ggagccgcct tttccgctgg
      841 gtgtcactcg ggggtgggga agatggccca ttcaaaagcg ccgcgagggg gcccggccag
      901 tgcccttcag tgagcgctcg caagaggacg gcagaggccc ggcagctcgg agctccggga
      961 ccttgtggcg catcaggacg cggctgtccc tctgccggga cccagagccg ccgccgccgc
     1021 tctgcctcct gcgtgttagc ctcctctgcg cgctccgggc aggcggccgt gggagccgct
     1081 ggggcgagga cggcgcgagg ctgctgctgc tgcccccggc ccgcgcggct ggaaacggag
     1141 aggccgagcc aagcggcggc ccctcttatg ctgggaggat gctggagagt agcggctgca
     1201 aagcgctgaa ggagggcgtg ctggagaagc gcagcgacgg gttgttgcag ctctggaaga
     1261 aaaagtgttg catcctcacc gaggaagggc tgctgcttat cccgcccaag cagctgcaac
     1321 accagcagca gcagcagcaa cagcagcagc agcagcagca acaacagccc gggcaggggc
     1381 cggccgagcc gtcccaaccc agtggccccg ctgtcgccag cctcgagccg ccggtcaagc
     1441 tcaaggaact gcacttctcc aacatgaaga ccgtggactg tgtggagcgc aagggcaagt
     1501 acatgtactt cactgtggtg atggcagagg gcaaggagat cgactttcgg tgcccgcaag
     1561 accagggctg gaacgccgag atcacgctgc agatggtgca gtacaagaat cgtcaggcca
     1621 tcctggcggt caaatccacg cggcagaagc agcagcacct ggtccagcag cagcccccct
     1681 cgcagccgca gccgcagccg cagctccagc cccaacccca gcctcagcct cagccgcaac
     1741 cccagcccca atcacaaccc cagcctcagc cccaacccaa gcctcagccc cagcagctcc
     1801 acccgtatcc gcatccacat ccacatccac actctcatcc tcactcgcac ccacaccctc
     1861 acccgcaccc gcatccgcac caaataccgc acccacaccc acagccgcac tcgcagccgc
     1921 acgggcaccg gcttctccgc agcacctcca actctgccgg atccgactac aaagaccatg
     1981 acggtgatta taaagatcat gacatcgact acaaggatga cgatgacaag tagtgagggc
     2041 ccgatatctc gacaatcaac ctctggatta caaaatttgt gaaagattga ctggtattct
     2101 taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc
     2161 tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct
     2221 ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga
     2281 cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc
     2341 tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac
     2401 aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaagc tgacgtcctt
     2461 tccatggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt
     2521 cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc
     2581 tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc
     2641 gcctggaaac gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc
     2701 gctatgacgg caataaaaag acagaataaa acgcacgggt gttgggtcgt ttgttcataa
     2761 acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg
     2821 ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc
     2881 agggctcgca gccaacgtcg gggcggcagg ccctgccata gcagatctgc gcagctgggg
     2941 ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     3001 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     3061 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc ggggcatccc
     3121 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     3181 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     3241 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     3301 ctattctttt gatttataag ggattttggg gatttcggcc tattggttaa aaaatgagct
     3361 gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga
     3421 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     3481 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     3541 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     3601 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     3661 gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     3721 ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca agagacagga
     3781 tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg
     3841 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
     3901 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
     3961 gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
     4021 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
     4081 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
     4141 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
     4201 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag
     4261 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag
     4321 gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat
     4381 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg
     4441 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
     4501 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
     4561 ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgcg aaatgaccga
     4621 ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag
     4681 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
     4741 catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata
     4801 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     4861 tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag
     4921 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     4981 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     5041 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     5101 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     5161 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     5221 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     5281 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     5341 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     5401 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     5461 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     5521 ggcgctttct caatgctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     5581 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     5641 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     5701 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     5761 ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     5821 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     5881 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     5941 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     6001 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     6061 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     6121 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     6181 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     6241 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     6301 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     6361 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     6421 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     6481 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     6541 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     6601 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     6661 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     6721 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     6781 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     6841 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     6901 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     6961 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     7021 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     7081 gccacctgac gtcgacggat cgggagatct cccgatcccc tatggtcgac tctcagtaca
     7141 atctgctctg atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc
     7201 gctgagtagt gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc
     7261 atgaagaatc tgcttagg
//
