LOCUS       V025754                 7492 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V025754
VERSION     V025754
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        202..581
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        582..785
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        830..848
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             931..2931
                     /label="NEDD1(NM_001135175)"
                     /note="NEDD1(NM_001135175)"
                     /gene="NEDD1"
     CDS             2938..2964
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             2968..2994
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             2998..3024
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     polyA_signal    3059..3283
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3329..3757
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3771..4100
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4167..4961
                     /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    5135..5256
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5305..5321)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5329..5345
                     /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(5353..5383)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5398..5419
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5707..6292)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6463..7323)
                     /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(7324..7428)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
       61 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
      121 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg
      181 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta
      241 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      301 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      361 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      421 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
      481 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
      541 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      601 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      661 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
      721 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      781 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc
      841 actataggga gacccaagct ggctagcgtt taaacttaag cttggtaccg agctcggatc
      901 cactagtcca gtgtggtgga attcgccacc atgcatttta caggcgcagt catgcaggaa
      961 aacctcagat ttgcttcatc aggagatgat attaaaatat gggatgcttc atctatgaca
     1021 ttggtggata aattcaaccc acacacatca ccacatggaa tcagctcaat atgttggagc
     1081 agcaataata actttttagt aacagcatct tccagtggcg acaaaatagt tgtctcaagt
     1141 tgcaaatgta aacctgttcc acttttagag cttgctgaag ggcaaaagca gacatgtgtc
     1201 aatttaaatt ctacatctat gtatttggta agcggaggcc taaataacac tgttaatatt
     1261 tgggatttaa aatcaaaaag agttcatcga tctcttaagg atcataaaga tcaagtaact
     1321 tgtgtaacat acaattggaa tgattgctac attgcttctg gatctcttag tggtgaaatt
     1381 attttacaca gtgtaaccac taatttatct agtactcctt ttggccatgg tagtaaccag
     1441 tctgttcggc acttgaagta ctccttgttt aagaaatcac tactgggcag tgtttcggat
     1501 aatggaatag taactctctg ggatgtaaat agtcagagtc cataccataa ctttgacagt
     1561 gtacacaaag ctccagcgtc aggcatctgt ttttctcctg tcaatgaatt gctctttgta
     1621 accataggct tggataaaag aatcatcctc tatgacactt caagtaagaa gctagtgaaa
     1681 actttagtgg ctgacactcc tctaactgcg gtagatttca tgcctgatgg agccactttg
     1741 gctattggat cttcccgggg gaaaatatat caatatgatt taagaatgtt gaaatcacca
     1801 gttaagacca tcagtgctca caagacatct gtgcagtgta tagcatttca gtactccact
     1861 gttcttacta agtcaagttt aaataaaggc tgttcaaata agcccacaac agtgaacaaa
     1921 cgaagtgtta atgtgaatgc tgctagtgga ggagttcaga attccggaat tgtcagagaa
     1981 gcacctgcca cgtccattgc cacagttcta ccacaaccta tgacatcagc tatggggaaa
     2041 ggaacagttg ctgttcaaga aaaagcaggt ttgcctcgaa gcataaacac agacacttta
     2101 tctaaggaaa cagacagtgg aaaaaatcag gatttctcca gctttgatga tactgggaaa
     2161 agtagtttag gtgacatgtt ctcacctatc agagatgatg ctgtagttaa caagggaagt
     2221 gatgagtcca taggcaaagg agatggcttt gactttctac cgcagttgaa ctcagtgttt
     2281 cctccaagaa aaaatccagt aacttcaagt acttcagtat tgcattctag tcctcttaat
     2341 gtttttatgg gatctccagg gaaagaggaa aatgaaaacc gtgatctaac agctgagtct
     2401 aagaaaatat atatgggaaa acaggaatct aaagactcct tcaaacagtt agcaaagttg
     2461 gtcacatctg gtgctgaaag tggaaatcta aatacctctc catcatctaa ccaaacaaga
     2521 aattctgaga aatttgaaaa gccagagaat gaaattgaag cccagttgat atgtgaaccc
     2581 ccaatcaatg gatcctcaac tccaaatcca aagatagcat cttctgtcac tgctggagtt
     2641 gccagttcac tctcagaaaa aatagccgac agcattggaa ataaccggca aaatgcacca
     2701 ttgacttcca ttcaaattcg ttttattcag aacatgatac aggaaacgtt ggatgacttt
     2761 agagaagcat gccataggga cattgtgaat ttgcaagtgg agatgattaa acagtttcat
     2821 atgcaactga atgaaatgca ttctttgctg gaaagatact cagtgaatga aggtttagtg
     2881 gctgaaattg aaagactacg agaagaaaac aaaagattac gggcccactt tctcgagtac
     2941 ccatacgacg tacctgatta tgcaacttat ccttatgatg tacctgatta tgctagctat
     3001 ccttatgatg tcccagacta cgcatgaggg cccgtttaaa cccgctgatc agcctcgact
     3061 gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg
     3121 gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg
     3181 agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg
     3241 gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga
     3301 accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg
     3361 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     3421 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     3481 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     3541 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     3601 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     3661 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     3721 aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt
     3781 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     3841 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     3901 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     3961 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     4021 cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg
     4081 gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc
     4141 aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc
     4201 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct
     4261 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg
     4321 acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca
     4381 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc
     4441 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga
     4501 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc
     4561 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc
     4621 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg
     4681 ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct
     4741 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc
     4801 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc
     4861 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc
     4921 agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga
     4981 aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt
     5041 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
     5101 cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg
     5161 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     5221 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc
     5281 tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     5341 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     5401 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     5461 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     5521 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     5581 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     5641 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     5701 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     5761 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     5821 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     5881 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     5941 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     6001 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     6061 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     6121 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     6181 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     6241 cggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     6301 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     6361 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     6421 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     6481 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     6541 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     6601 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     6661 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     6721 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     6781 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     6841 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     6901 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     6961 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     7021 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     7081 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     7141 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     7201 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     7261 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     7321 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     7381 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     7441 aaaagtgcca cctgacgtcg acggatcggg agatctcccg atcccctatg gt
//
