LOCUS       V018303                 6837 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V018303
VERSION     V018303
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             907..2298
                     /label="MYCN(NM_005378)"
                     /note="MYCN(NM_005378)"
                     /gene="MYCN"
     CDS             2299..2325
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     promoter        complement(2389..2407)
                     /note="promoter for bacteriophage SP6 RNA polymerase"
                     /label="SP6 promoter"
     polyA_signal    2433..2657
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2703..3131
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3145..3475
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3542..4336
                     /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    4510..4631
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4680..4696)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4704..4720
                     /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(4728..4758)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4773..4794
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5082..5670)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5841..6701)
                     /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(6702..6806)
                     /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 gccaccatgc cgagctgctc cacgtccacc atgccgggca tgatctgcaa gaacccagac
      961 ctcgagtttg actcgctaca gccctgcttc tacccggacg aagatgactt ctacttcggc
     1021 ggccccgact cgaccccccc gggggaggac atctggaaga agtttgagct gctgcccacg
     1081 cccccgctgt cgcccagccg tggcttcgcg gagcacagct ccgagccccc gagctgggtc
     1141 acggagatgc tgcttgagaa cgagctgtgg ggcagcccgg ccgaggagga cgcgttcggc
     1201 ctggggggac tgggtggcct cacccccaac ccggtcatcc tccaggactg catgtggagc
     1261 ggcttctccg cccgcgagaa gctggagcgc gccgtgagcg agaagctgca gcacggccgc
     1321 gggccgccaa ccgccggttc caccgcccag tccccgggag ccggcgccgc cagccctgcg
     1381 ggtcgcgggc acggcggggc tgcgggagcc ggccgcgccg gggccgccct gcccgccgag
     1441 ctcgcccacc cggccgccga gtgcgtggat cccgccgtgg tcttcccctt tcccgtgaac
     1501 aagcgcgagc cagcgcccgt gcccgcagcc ccggccagtg ccccggcggc gggccctgcg
     1561 gtcgcctcgg gggcgggtat tgccgcccca gccggggccc cgggggtcgc ccctccgcgc
     1621 ccaggcggcc gccagaccag cggcggcgac cacaaggccc tcagtacctc cggagaggac
     1681 accctgagcg attcagatga tgaagatgat gaagaggaag atgaagagga agaaatcgac
     1741 gtggtcactg tggagaagcg gcgttcctcc tccaacacca aggctgtcac cacattcacc
     1801 atcactgtgc gtcccaagaa cgcagccctg ggtcccggga gggctcagtc cagcgagctg
     1861 atcctcaaac gatgccttcc catccaccag cagcacaact atgccgcccc ctctccctac
     1921 gtggagagtg aggatgcacc cccacagaag aagataaaga gcgaggcgtc cccacgtccg
     1981 ctcaagagtg tcatcccccc aaaggctaag agcttgagcc cccgaaactc tgactcggag
     2041 gacagtgagc gtcgcagaaa ccacaacatc ctggagcgcc agcgccgcaa cgaccttcgg
     2101 tccagctttc tcacgctcag ggaccacgtg ccggagttgg taaagaatga gaaggccgcc
     2161 aaggtggtca ttttgaaaaa ggccactgag tatgtccact ccctccaggc cgaggagcac
     2221 cagcttttgc tggaaaagga aaaattgcag gcaagacagc agcagttgct aaagaaaatt
     2281 gaacacgctc ggacttgcta cccatacgac gtcccagact acgcttagga attctgcaga
     2341 tatccatcac actggcggcc gctcgagcat gcatctagag ggccctattc tatagtgtca
     2401 cctaaatgct agagctcgct gatcagcctc gactgtgcct tctagttgcc agccatctgt
     2461 tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc
     2521 ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg
     2581 tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga
     2641 tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc
     2701 ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     2761 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     2821 cacgttcgcc ggctttcccc gtcaagctct aaatcggggc atccctttag ggttccgatt
     2881 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     2941 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     3001 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     3061 ataagggatt ttggggattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     3121 taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
     3181 ccaggcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
     3241 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     3301 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     3361 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc
     3421 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct
     3481 cccgggagct tgtatatcca ttttcggatc tgatcaagag acaggatgag gatcgtttcg
     3541 catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt
     3601 cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc
     3661 agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact
     3721 gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt
     3781 gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca
     3841 ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat
     3901 gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg
     3961 catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga
     4021 agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga
     4081 cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa
     4141 tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga
     4201 catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt
     4261 cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct
     4321 tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac
     4381 ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc
     4441 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc
     4501 gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     4561 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     4621 aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg
     4681 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     4741 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     4801 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     4861 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     4921 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     4981 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     5041 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     5101 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     5161 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     5221 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcaatgc
     5281 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     5341 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     5401 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     5461 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     5521 aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     5581 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     5641 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     5701 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     5761 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     5821 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     5881 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     5941 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     6001 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     6061 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     6121 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     6181 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     6241 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     6301 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     6361 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     6421 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     6481 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     6541 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     6601 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     6661 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     6721 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     6781 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtc
//
