LOCUS       V018211                 7387 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V018211
VERSION     V018211
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             921..947
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             951..2858
                     /label="TP73(NM_005427)"
                     /note="TP73(NM_005427)"
                     /gene="TP73"
     promoter        complement(2939..2957)
                     /note="promoter for bacteriophage SP6 RNA polymerase"
                     /label="SP6 promoter"
     polyA_signal    2983..3207
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3253..3681
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3695..4025
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4092..4886
                     /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    5060..5181
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5230..5246)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5254..5270
                     /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(5278..5308)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5323..5344
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5632..6220)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6391..7251)
                     /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(7252..7356)
                     /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 ggagacccaa gcttggtacc
      901 gagctcggat ctctatggct tacccatacg atgttccaga ttacgctagc atggcccagt
      961 ccaccgccac ctcccctgat gggggcacca cgtttgagca cctctggagc tctctggaac
     1021 cagacagcac ctacttcgac cttccccagt caagccgggg gaataatgag gtggtgggcg
     1081 gaacggattc cagcatggac gtcttccacc tggagggcat gactacatct gtcatggccc
     1141 agttcaatct gctgagcagc accatggacc agatgagcag ccgcgcggcc tcggccagcc
     1201 cctacacccc agagcacgcc gccagcgtgc ccacccactc gccctacgca caacccagct
     1261 ccaccttcga caccatgtcg ccggcgcctg tcatcccctc caacaccgac taccccggac
     1321 cccaccactt tgaggtcact ttccagcagt ccagcacggc caagtcagcc acctggacgt
     1381 actccccgct cttgaagaaa ctctactgcc agatcgccaa gacatgcccc atccagatca
     1441 aggtgtccac cccgccaccc ccaggcactg ccatccgggc catgcctgtt tacaagaaag
     1501 cggagcacgt gaccgacgtc gtgaaacgct gccccaacca cgagctcggg agggacttca
     1561 acgaaggaca gtctgctcca gccagccacc tcatccgcgt ggaaggcaat aatctctcgc
     1621 agtatgtgga tgaccctgtc accggcaggc agagcgtcgt ggtgccctat gagccaccac
     1681 aggtggggac ggaattcacc accatcctgt acaacttcat gtgtaacagc agctgtgtag
     1741 ggggcatgaa ccggcggccc atcctcatca tcatcaccct ggagatgcgg gatgggcagg
     1801 tgctgggccg ccggtccttt gagggccgca tctgcgcctg tcctggccgc gaccgaaaag
     1861 ctgatgagga ccactaccgg gagcagcagg ccctgaacga gagctccgcc aagaacgggg
     1921 ccgccagcaa gcgtgccttc aagcagagcc cccctgccgt ccccgccctt ggtgccggtg
     1981 tgaagaagcg gcggcatgga gacgaggaca cgtactacct tcaggtgcga ggccgggaga
     2041 actttgagat cctgatgaag ctgaaagaga gcctggagct gatggagttg gtgccgcagc
     2101 cactggtgga ctcctatcgg cagcagcagc agctcctaca gaggccgagt cacctacagc
     2161 ccccgtccta cgggccggtc ctctcgccca tgaacaaggt gcacgggggc atgaacaagc
     2221 tgccctccgt caaccagctg gtgggccagc ctcccccgca cagttcggca gctacaccca
     2281 acctggggcc cgtgggcccc gggatgctca acaaccatgg ccacgcagtg ccagccaacg
     2341 gcgagatgag cagcagccac agcgcccagt ccatggtctc ggggtcccac tgcactccgc
     2401 caccccccta ccacgccgac cccagcctcg tcagtttttt aacaggattg gggtgtccaa
     2461 actgcatcga gtatttcacc tcccaagggt tacagagcat ttaccacctg cagaacctga
     2521 ccattgagga cctgggggcc ctgaagatcc ccgagcagta ccgcatgacc atctggcggg
     2581 gcctgcagga cctgaagcag ggccacgact acagcaccgc gcagcagctg ctccgctcta
     2641 gcaacgcggc caccatctcc atcggcggct caggggaact gcagcgccag cgggtcatgg
     2701 aggccgtgca cttccgcgtg cgccacacca tcaccatccc caaccgcggc ggcccaggcg
     2761 gcggccctga cgagtgggcg gacttcggct tcgacctgcc cgactgcaag gcccgcaagc
     2821 agcccatcaa ggaggagttc acggaggccg agatccactg agggccacgc ctgaagccga
     2881 attctgcaga tatccatcac actggcggcc gctcgagcat gcatctagag ggccctattc
     2941 tatagtgtca cctaaatgct agagctcgct gatcagcctc gactgtgcct tctagttgcc
     3001 agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca
     3061 ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta
     3121 ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc
     3181 atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta
     3241 gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     3301 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     3361 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggc atccctttag
     3421 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
     3481 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     3541 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
     3601 cttttgattt ataagggatt ttggggattt cggcctattg gttaaaaaat gagctgattt
     3661 aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc
     3721 cccaggctcc ccaggcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     3781 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     3841 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     3901 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     3961 cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     4021 gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag acaggatgag
     4081 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg
     4141 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
     4201 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc
     4261 tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt
     4321 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag
     4381 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg
     4441 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag
     4501 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
     4561 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc
     4621 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca
     4681 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc
     4741 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg
     4801 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct
     4861 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc
     4921 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg
     4981 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     5041 ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa
     5101 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     5161 caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc
     5221 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     5281 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     5341 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     5401 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     5461 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     5521 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     5581 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     5641 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     5701 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     5761 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     5821 ttctcaatgc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     5881 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     5941 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     6001 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     6061 ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     6121 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     6181 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     6241 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     6301 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     6361 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     6421 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     6481 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     6541 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     6601 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     6661 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     6721 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     6781 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     6841 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     6901 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     6961 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     7021 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     7081 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     7141 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     7201 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     7261 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     7321 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     7381 tgacgtc
//
