LOCUS       V030740                 8473 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V030740
VERSION     V030740
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"
     CDS             706..3129
                     /label="MCM3(NM_002388)"
                     /note="MCM3(NM_002388)"
                     /gene="MCM3"
     CDS             3136..3225
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     misc_feature    3249..3837
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3903..3951
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4153..4581
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4595..4924
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4991..5785
                     /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    5961..6082
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6131..6147)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6155..6171
                     /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(6179..6209)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6224..6245
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6533..7121)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7292..8152)
                     /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(8153..8257)
                     /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 gctctggcta gcgtttaaac ttaagcttgg
      661 taccgagctc ggatccacta gtccagtgtg gtggaattcg ccaccatggc gggtaccgtg
      721 gtgctggacg atgtggagct gcgggaggct cagagagatt acctggactt cctggacgac
      781 gaggaagacc agggaattta tcagagcaaa gttcgggagc tgatcagtga caaccaatac
      841 cggctgattg tcaatgtgaa tgacctgcgc aggaaaaacg agaagagggc taaccggctt
      901 ctgaacaatg cctttgagga gctggttgcc ttccagcggg ccttaaagga ttttgtggcc
      961 tccattgatg ctacctatgc caagcagtat gaggagttct acgtaggact ggaaggcagc
     1021 tttggctcca agcacgtctc cccgcggact cttacctcct gcttcctcag ctgtgtggtc
     1081 tgtgtggagg gcattgtcac taaatgttct ctagttcgtc ccaaagtcgt ccgcagtgtc
     1141 cactactgtc ctgctactaa gaagaccata gagcgacgtt attctgatct caccaccctg
     1201 gtggcctttc cctccagctc tgtctatcct accaaggatg aggagaacaa tccccttgag
     1261 acagaatatg gcctttctgt ctacaaggat caccagacca tcaccatcca ggagatgccg
     1321 gagaaggccc cagccggcca gctcccccgc tctgtggacg tcattctgga tgatgacttg
     1381 gtggataaag cgaagcctgg tgaccgggtt caggtggtgg gaacctaccg ttgccttcct
     1441 ggaaagaagg gaggctacac ctctgggacc ttcaggactg tcctgattgc ctgtaatgtt
     1501 aagcagatga gcaaggatgc tcagccctct ttctctgctg aggatatagc caagatcaag
     1561 aagttcagta aaacccgatc caaggatatc tttgaccagc tggccaagtc attggcccca
     1621 agtatccatg ggcatgacta tgtcaagaaa gcaatcctct gcttgctctt gggaggggtg
     1681 gaacgagacc tagaaaatgg cagccacatc cgtggggaca tcaatattct tctaatagga
     1741 gacccatccg ttgccaagtc tcagcttctg cggtatgtgc tttgcactgc accccgagct
     1801 atccccacca ctggccgggg ctcctctgga gtgggtctga cggctgctgt caccacagac
     1861 caggaaacag gagagcgccg tctggaagca ggggccatgg tcctggctga ccgaggcgtg
     1921 gtttgcattg atgaatttga caaaatgtct gacatggatc gcacagccat ccatgaagtg
     1981 atggagcagg gtcgagtgac cattgccaag gctggcatcc atgctcggct gaatgcccgc
     2041 tgcagtgttt tggcagctgc caaccctgtc tacggcaggt atgaccagta taagactcca
     2101 atggagaaca ttgggctaca ggactcactg ctgtcacgat ttgacttgct cttcatcatg
     2161 ctggatcaga tggatcctga gcaggatcgg gagatctcag accatgtcct tcggatgcac
     2221 cgttacagag cacctgggga gcaggatggc gatgctatgc ccttgggtag tgctgtggat
     2281 atcctggcca cagatgatcc caactttagc caggaagatc agcaggacac ccagatttat
     2341 gagaagcatg acaaccttct acatgggacc aagaagaaaa aggagaagat ggtgagtgca
     2401 gcattcatga agaagtacat ccatgtggcc aaaatcatca agcctgtcct gacacaggag
     2461 tcggccacct acattgcaga agagtattca cgcctgcgca gccaggatag catgagctca
     2521 gacaccgcca ggacatctcc agttacagcc cgaacactgg aaactctgat tcgactggcc
     2581 acagcccatg cgaaggcccg catgagcaag actgtggacc tgcaggatgc agaggaagct
     2641 gtggagttgg tccagtatgc ttactttaag aaggttctgg agaaggagaa gaaacgtaag
     2701 aagcgaagtg aggatgaatc agagacagaa gatgaagagg agaaaagcca agaggaccag
     2761 gagcagaaga ggaagagaag gaagactcgc cagccagatg ccaaagatgg ggattcatac
     2821 gacccctatg acttcagtga cacagaggag gaaatgcctc aagtacacac tccaaagacg
     2881 gcagactcac aggagaccaa ggaatcccag aaagtggagt tgagtgaatc caggttgaag
     2941 gcattcaagg tggccctctt ggatgtgttc cgggaagctc atgcgcagtc aatcggcatg
     3001 aatcgcctca cagaatccat caaccgggac agcgaagagc ccttctcttc agttgagatc
     3061 caggctgctc tgagcaagat gcaggatgac aatcaggtca tggtgtctga gggcatcatc
     3121 ttcctcatcc tcgaggaaca aaaactaata tcagaggaag atttggaaca gaaactaatc
     3181 agtgaagaag atttagagca aaagttaatt tccgaggagg acttatagtg agggcccgat
     3241 atctcgacaa tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact
     3301 atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg
     3361 cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg
     3421 aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa
     3481 cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc
     3541 ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg
     3601 ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaagctgacg tcctttccat
     3661 ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt
     3721 cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc
     3781 cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcctg
     3841 gaaacggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa cccgcgctat
     3901 gacggcaata aaaagacaga ataaaacgca cgggtgttgg gtcgtttgtt cataaacgcg
     3961 gggttcggtc ccagggctgg cactctgtcg ataccccacc gagaccccat tggggccaat
     4021 acgcccgcgt ttcttccttt tccccacccc accccccaag ttcgggtgaa ggcccagggc
     4081 tcgcagccaa cgtcggggcg gcaggccctg ccatagcaga tctgcgcagc tggggctcta
     4141 gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     4201 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     4261 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggc atccctttag
     4321 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
     4381 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     4441 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
     4501 cttttgattt ataagggatt ttggggattt cggcctattg gttaaaaaat gagctgattt
     4561 aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc
     4621 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
     4681 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     4741 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     4801 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     4861 ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     4921 caaaaagctc ccgggagctt gtatatccat tttcggatct gatcaagaga caggatgagg
     4981 atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga
     5041 gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt
     5101 ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct
     5161 gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg
     5221 cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt
     5281 gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc
     5341 tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc
     5401 gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga
     5461 tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg
     5521 catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat
     5581 ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg
     5641 ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc
     5701 tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta
     5761 tcgccttctt gacgagttct tctgagcggg actctggggt tcgcgaaatg accgaccaag
     5821 cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg
     5881 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc
     5941 tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca
     6001 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     6061 ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg
     6121 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     6181 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     6241 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     6301 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     6361 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     6421 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     6481 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     6541 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     6601 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     6661 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     6721 tttctcaatg ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     6781 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     6841 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     6901 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     6961 gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     7021 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     7081 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     7141 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     7201 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     7261 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     7321 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     7381 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     7441 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     7501 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     7561 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     7621 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     7681 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     7741 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     7801 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     7861 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     7921 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     7981 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     8041 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     8101 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     8161 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     8221 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     8281 ctgacgtcga cggatcggga gatctcccga tcccctatgg tcgactctca gtacaatctg
     8341 ctctgatgcc gcatagttaa gccagtatct gctccctgct tgtgtgttgg aggtcgctga
     8401 gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc ttgaccgaca attgcatgaa
     8461 gaatctgctt agg
//
