LOCUS       V020820                 6591 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V020820
VERSION     V020820
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        235..614
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        615..818
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        863..881
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             898..924
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             934..2115
                     /codon_start=1
                     /gene="Homo sapiens TP53"
                     /note="responds to cellular stresses to regulate the
                     expression of target genes"
                     /product="human tumor suppressor protein that is often
                     mutated in cancers"
                     /transl_table=1
                     /translation="MEEPQSDPSVEPPLSQETFSDLWKLLPENNVLSPLPSQAMDDLML
                     SPDDIEQWFTEDPGPDEAPRMPEAAPPVAPAPAAPTPAAPAPAPSWPLSSSVPSQKTYQ
                     GSYGFRLGFLHSGTAKSVTCTYSPALNKMFCQLAKTCPVQLWVDSTPPPGTRVRAMAIY
                     KQSQHMTEVVRRCPHHERCSDSDGLAPPQHLIRVEGNLRVEYLDDRNTFRHSVVVPYEP
                     PEVGSDCTTIHYNYMCNSSCMGGMNWRPILTIITLEDSSGNLLGRNSFEVRVCACPGRD
                     RRTEEENLRKKGEPHHELPPGSTKRALPNNTSSSPQPKKKPLDGEYFTLQIRGRERFEM
                     FRELNEALELKDAQAGKEPGGSRAHSSHLKSKKGQSTSRHKKLMFKTEGPDSD*"
                     /label="p53"
     promoter        complement(2143..2161)
                     /note="promoter for bacteriophage SP6 RNA polymerase"
                     /label="SP6 promoter"
     polyA_signal    2187..2411
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2457..2885
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        2899..3229
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3296..4090
                     /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    4264..4385
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4434..4450)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4458..4474
                     /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(4482..4512)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4527..4548
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4836..5424)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5595..6455)
                     /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(6456..6560)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggac tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccgc caccatgtac
      901 ccatacgacg tcccagacta cgctggaagc ttcatggagg agccgcagtc agatcctagc
      961 gtcgagcccc ctctgagtca ggaaacattt tcagacctat ggaaactact tcctgaaaac
     1021 aacgttctgt cccccttgcc gtcccaagca atggatgatt tgatgctgtc cccggacgat
     1081 attgaacaat ggttcactga agacccaggt ccagatgaag ctcccagaat gccagaggct
     1141 gctccccccg tggcccctgc accagcagct cctacaccgg cggcccctgc accagccccc
     1201 tcctggcccc tgtcatcttc tgtcccttcc cagaaaacct accagggcag ctacggtttc
     1261 cgtctgggct tcttgcattc tgggacagcc aagtctgtga cttgcacgta ctcccctgcc
     1321 ctcaacaaga tgttttgcca actggccaag acctgccctg tgcagctgtg ggttgattcc
     1381 acacccccgc ccggcacccg cgtccgcgcc atggccatct acaagcagtc acagcacatg
     1441 acggaggttg tgaggcgctg cccccaccat gagcgctgct cagatagcga tggtctggcc
     1501 cctcctcagc atcttatccg agtggaagga aatttgcgtg tggagtattt ggatgacaga
     1561 aacacttttc gacatagtgt ggtggtgccc tatgagccgc ctgaggttgg ctctgactgt
     1621 accaccatcc actacaacta catgtgtaac agttcctgca tgggcggcat gaactggagg
     1681 cccatcctca ccatcatcac actggaagac tccagtggta atctactggg acggaacagc
     1741 tttgaggtgc gtgtttgtgc ctgtcctggg agagaccggc gcacagagga agagaatctc
     1801 cgcaagaaag gggagcctca ccacgagctg cccccaggga gcactaagcg agcactgccc
     1861 aacaacacca gctcctctcc ccagccaaag aagaaaccac tggatggaga atatttcacc
     1921 cttcagatcc gtgggcgtga gcgcttcgag atgttccgag agctgaatga ggccttggaa
     1981 ctcaaggatg cccaggctgg gaaggagcca ggggggagca gggctcactc cagccacctg
     2041 aagtccaaaa agggtcagtc tacctcccgc cataaaaaac tcatgttcaa gacagaaggg
     2101 cctgactcag actgactcga gcatgcatct agagggccct attctatagt gtcacctaaa
     2161 tgctagagct cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg
     2221 cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata
     2281 aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt
     2341 ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt
     2401 gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc
     2461 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     2521 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     2581 cgccggcttt ccccgtcaag ctctaaatcg gggcatccct ttagggttcc gatttagtgc
     2641 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
     2701 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     2761 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     2821 gattttgggg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     2881 gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccaggc
     2941 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc
     3001 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt
     3061 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     3121 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct
     3181 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg
     3241 agcttgtata tccattttcg gatctgatca agagacagga tgaggatcgt ttcgcatgat
     3301 tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta
     3361 tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca
     3421 ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga
     3481 cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga
     3541 cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct
     3601 cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg
     3661 gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga
     3721 gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca
     3781 tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga
     3841 ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg
     3901 cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc
     3961 gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt
     4021 gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga
     4081 gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca
     4141 tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc
     4201 cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac
     4261 cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     4321 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     4381 tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag
     4441 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     4501 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     4561 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     4621 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     4681 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     4741 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     4801 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     4861 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     4921 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     4981 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca atgctcacgc
     5041 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     5101 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     5161 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     5221 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca
     5281 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     5341 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     5401 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     5461 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     5521 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     5581 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     5641 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     5701 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     5761 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     5821 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     5881 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     5941 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     6001 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     6061 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     6121 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     6181 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     6241 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     6301 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     6361 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     6421 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     6481 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     6541 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt c
//
