LOCUS       V022805                 7699 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V022805
VERSION     V022805
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..2358
                     /label="RELA(NM_021975)"
                     /note="RELA(NM_021975)"
                     /gene="RELA"
     CDS             2365..2391
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             2395..2421
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             2425..2451
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     misc_feature    2475..3063
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3129..3177
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3379..3807
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3821..4150
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4217..5011
                     /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    5187..5308
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5357..5373)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5381..5397
                     /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(5405..5435)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5450..5471
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5759..6347)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6518..7378)
                     /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(7379..7483)
                     /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 ccaccatgga cgaactgttc
      721 cccctcatct tcccggcaga gccagcccag gcctctggcc cctatgtgga gatcattgag
      781 cagcccaagc agcggggcat gcgcttccgc tacaagtgcg aggggcgctc cgcgggcagc
      841 atcccaggcg agaggagcac agataccacc aagacccacc ccaccatcaa gatcaatggc
      901 tacacaggac cagggacagt gcgcatctcc ctggtcacca aggaccctcc tcaccggcct
      961 cacccccacg agcttgtagg aaaggactgc cgggatggct tctatgaggc tgagctctgc
     1021 ccggaccgct gcatccacag tttccagaac ctgggaatcc agtgtgtgaa gaagcgggac
     1081 ctggagcagg ctatcagtca gcgcatccag accaacaaca accccttcca agttcctata
     1141 gaagagcagc gtggggacta cgacctgaat gctgtgcggc tctgcttcca ggtgacagtg
     1201 cgggacccat caggcaggcc cctccgcctg ccgcctgtcc tttctcatcc catctttgac
     1261 aatcgtgccc ccaacactgc cgagctcaag atctgccgag tgaaccgaaa ctctggcagc
     1321 tgcctcggtg gggatgagat cttcctactg tgtgacaagg tgcagaaaga ggacattgag
     1381 gtgtatttca cgggaccagg ctgggaggcc cgaggctcct tttcgcaagc tgatgtgcac
     1441 cgacaagtgg ccattgtgtt ccggacccct ccctacgcag accccagcct gcaggctcct
     1501 gtgcgtgtct ccatgcagct gcggcggcct tccgaccggg agctcagtga gcccatggaa
     1561 ttccagtacc tgccagatac agacgatcgt caccggattg aggagaaacg taaaaggaca
     1621 tatgagacct tcaagagcat catgaagaag agtcctttca gcggacccac cgacccccgg
     1681 cctccacctc gacgcattgc tgtgccttcc cgcagctcag cttctgtccc caagccagca
     1741 ccccagccct atccctttac gtcatccctg agcaccatca actatgatga gtttcccacc
     1801 atggtgtttc cttctgggca gatcagccag gcctcggcct tggccccggc ccctccccaa
     1861 gtcctgcccc aggctccagc ccctgcccct gctccagcca tggtatcagc tctggcccag
     1921 gccccagccc ctgtcccagt cctagcccca ggccctcctc aggctgtggc cccacctgcc
     1981 cccaagccca cccaggctgg ggaaggaacg ctgtcagagg ccctgctgca gctgcagttt
     2041 gatgatgaag acctgggggc cttgcttggc aacagcacag acccagctgt gttcacagac
     2101 ctggcatccg tcgacaactc cgagtttcag cagctgctga accagggcat acctgtggcc
     2161 ccccacacaa ctgagcccat gctgatggag taccctgagg ctataactcg cctagtgaca
     2221 ggggcccaga ggccccccga cccagctcct gctccactgg gggccccggg gctccccaat
     2281 ggcctccttt caggagatga agacttctcc tccattgcgg acatggactt ctcagccctg
     2341 ctgagtcaga tcagctccct cgagtaccca tacgacgtac ctgattatgc aacttatcct
     2401 tatgatgtac ctgattatgc tagctatcct tatgatgtcc cagactacgc atagtgaggg
     2461 cccgatatct cgacaatcaa cctctggatt acaaaatttg tgaaagattg actggtattc
     2521 ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg
     2581 ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc
     2641 tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg
     2701 acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg
     2761 ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga
     2821 caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaag ctgacgtcct
     2881 ttccatggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg
     2941 tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc
     3001 ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc
     3061 cgcctggaaa cgggggaggc taactgaaac acggaaggag acaataccgg aaggaacccg
     3121 cgctatgacg gcaataaaaa gacagaataa aacgcacggg tgttgggtcg tttgttcata
     3181 aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga ccccattggg
     3241 gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg ggtgaaggcc
     3301 cagggctcgc agccaacgtc ggggcggcag gccctgccat agcagatctg cgcagctggg
     3361 gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     3421 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     3481 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cggggcatcc
     3541 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
     3601 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     3661 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
     3721 tctattcttt tgatttataa gggattttgg ggatttcggc ctattggtta aaaaatgagc
     3781 tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg
     3841 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     3901 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     3961 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
     4021 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     4081 ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     4141 cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc aagagacagg
     4201 atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg
     4261 ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc
     4321 cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg
     4381 tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt
     4441 tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg
     4501 cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat
     4561 catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca
     4621 ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca
     4681 ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa
     4741 ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa
     4801 tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc
     4861 ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga
     4921 atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc
     4981 cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcgc gaaatgaccg
     5041 accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa
     5101 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
     5161 tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat
     5221 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     5281 gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga
     5341 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     5401 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     5461 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     5521 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     5581 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     5641 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     5701 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     5761 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     5821 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     5881 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     5941 tggcgctttc tcaatgctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     6001 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     6061 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     6121 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     6181 actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct
     6241 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     6301 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     6361 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     6421 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     6481 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     6541 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     6601 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     6661 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     6721 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     6781 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     6841 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     6901 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     6961 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     7021 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     7081 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     7141 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     7201 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     7261 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     7321 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     7381 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     7441 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     7501 tgccacctga cgtcgacgga tcgggagatc tcccgatccc ctatggtcga ctctcagtac
     7561 aatctgctct gatgccgcat agttaagcca gtatctgctc cctgcttgtg tgttggaggt
     7621 cgctgagtag tgcgcgagca aaatttaagc tacaacaagg caaggcttga ccgacaattg
     7681 catgaagaat ctgcttagg
//
