LOCUS       V020115                 7019 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V020115
VERSION     V020115
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"
     regulatory      677..686
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             701..1678
                     /label="ZFP36(NM_003407)"
                     /note="ZFP36(NM_003407)"
                     /gene="ZFP36"
     CDS             1685..1711
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             1715..1741
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             1745..1771
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     misc_feature    1795..2383
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2449..2497
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      2699..3127
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3141..3470
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3537..4331
                     /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    4507..4628
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4677..4693)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4701..4717
                     /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(4725..4755)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4770..4791
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5079..5667)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5838..6698)
                     /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(6699..6803)
                     /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 ggatccgcca ccatggccaa ccgttacacc atggatctga ctgccatcta
      721 cgagagcctc ctgtcgctga gccctgacgt gcccgtgcca tccgaccatg gagggactga
      781 gtccagccca ggctggggct cctcgggacc ctggagcctg agcccctccg actccagccc
      841 gtctggggtc acctcccgcc tgcctggccg ctccaccagc ctagtggagg gccgcagctg
      901 tggctgggtg cccccacccc ctggcttcgc accgctggct ccccgcctgg gccctgagct
      961 gtcaccctca cccacttcgc ccactgcaac ctccaccacc ccctcgcgct acaagactga
     1021 gctatgtcgg accttctcag agagtgggcg ctgccgctac ggggccaagt gccagtttgc
     1081 ccatggcctg ggcgagctgc gccaggccaa tcgccacccc aaatacaaga cggaactctg
     1141 tcacaagttc tacctccagg gccgctgccc ctacggctct cgctgccact tcatccacaa
     1201 ccctagcgaa gacctggcgg ccccgggcca ccctcctgtg cttcgccaga gcatcagctt
     1261 ctccggcctg ccctctggcc gccggacctc accaccacca ccaggcctgg ccggcccttc
     1321 cctgtcctcc agctccttct cgccctccag ctccccacca ccacctgggg accttccact
     1381 gtcaccctct gccttctctg ctgcccctgg cacccccctg gctcgaagag accccacccc
     1441 agtctgttgc ccctcctgcc gaagggccac tcctatcagc gtctgggggc ccttgggtgg
     1501 cctggttcgg accccctctg tacagtccct gggatccgac cctgatgaat atgccagcag
     1561 cggcagcagc ctggggggct ctgactctcc cgtcttcgag gcgggagttt ttgcaccacc
     1621 ccagcccgtg gcagcccccc ggcgactccc catcttcaat cgcatctctg tttctgagct
     1681 cgagtaccca tacgacgtac ctgattatgc aacttatcct tatgatgtac ctgattatgc
     1741 tagctatcct tatgatgtcc cagactacgc atagtgaggg cccgatatct cgacaatcaa
     1801 cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt
     1861 acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct
     1921 ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc
     1981 gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg
     2041 ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc
     2101 acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc
     2161 actgacaatt ccgtggtgtt gtcggggaag ctgacgtcct ttccatggct gctcgcctgt
     2221 gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca
     2281 gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt
     2341 cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcctggaaa cgggggaggc
     2401 taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa
     2461 gacagaataa aacgcacggg tgttgggtcg tttgttcata aacgcggggt tcggtcccag
     2521 ggctggcact ctgtcgatac cccaccgaga ccccattggg gccaatacgc ccgcgtttct
     2581 tccttttccc caccccaccc cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc
     2641 ggggcggcag gccctgccat agcagatctg cgcagctggg gctctagggg gtatccccac
     2701 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     2761 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     2821 ttcgccggct ttccccgtca agctctaaat cggggcatcc ctttagggtt ccgatttagt
     2881 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     2941 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     3001 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     3061 gggattttgg ggatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     3121 gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     3181 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     3241 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     3301 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     3361 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc
     3421 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg
     3481 gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg tttcgcatga
     3541 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
     3601 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
     3661 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg
     3721 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
     3781 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
     3841 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
     3901 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
     3961 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
     4021 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg
     4081 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
     4141 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
     4201 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
     4261 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
     4321 agttcttctg agcgggactc tggggttcgc gaaatgaccg accaagcgac gcccaacctg
     4381 ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt
     4441 ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc
     4501 caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     4561 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     4621 gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca
     4681 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     4741 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     4801 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     4861 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     4921 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     4981 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     5041 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     5101 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     5161 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     5221 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcaatgctca
     5281 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     5341 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     5401 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     5461 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg
     5521 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     5581 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     5641 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     5701 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     5761 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     5821 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     5881 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     5941 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     6001 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     6061 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     6121 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     6181 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     6241 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     6301 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     6361 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     6421 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     6481 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     6541 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     6601 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     6661 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     6721 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     6781 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacgga
     6841 tcgggagatc tcccgatccc ctatggtcga ctctcagtac aatctgctct gatgccgcat
     6901 agttaagcca gtatctgctc cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca
     6961 aaatttaagc tacaacaagg caaggcttga ccgacaattg catgaagaat ctgcttagg
//
