LOCUS       V023321                 7372 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V023321
VERSION     V023321
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"
     regulatory      913..922
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             922..987
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     CDS             1009..2892
                     /label="PIAS3(NM_006099)"
                     /note="PIAS3(NM_006099)"
                     /gene="PIAS3"
     polyA_signal    2972..3196
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3242..3670
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3684..4013
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4080..4874
                     /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    5048..5169
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5218..5234)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5242..5258
                     /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(5266..5296)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5311..5332
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5620..6205)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6376..7236)
                     /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(7237..7341)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct 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 atgggtggag 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 gctggctagc
      901 gtttaaactt aagccaccat ggactacaaa gaccatgacg gtgattataa agatcatgac
      961 atcgactaca aggatgacga tgacaagctt ggtaccgagc tcggatccat ggcggagctg
     1021 ggcgaattaa agcacatggt gatgagtttc cgggtgtctg agctccaggt gcttcttggc
     1081 tttgctggcc ggaacaagag tggacggaag cacgagctcc tggccaaggc tctgcacctc
     1141 ctgaagtcca gctgtgcccc tagtgtccag atgaagatca aagagcttta ccgacgacgc
     1201 tttccccgga agaccctggg gccctctgat ctctcccttc tctctttgcc ccctggcacc
     1261 tctcctgtag gctcccctgg tcctctagct cccattcccc caacgctgtt ggcccctggc
     1321 accctgctgg gccccaagcg tgaggtggac atgcaccccc ctctgcccca gcctgtgcac
     1381 cctgatgtca ccatgaaacc attgcccttc tatgaagtct atggggagct catccggccc
     1441 accacccttg catccacttc tagccagcgg tttgaggaag cgcactttac ctttgccctc
     1501 acaccccagc aagtgcagca gattcttaca tccagagagg ttctgccagg agccaaatgt
     1561 gattatacca tacaggtgca gctaaggttc tgtctctgtg agaccagctg cccccaggaa
     1621 gattattttc cccccaacct ctttgtcaag gtcaatggga aactgtgccc cctgccgggt
     1681 taccttcccc caaccaagaa tggggccgag cccaagaggc ccagccgccc catcaacatc
     1741 acacccctgg ctcgactctc agccactgtt cccaacacca ttgtggtcaa ttggtcatct
     1801 gagttcggac ggaattactc cttgtctgtg tacctggtga ggcagttgac tgcaggaacc
     1861 cttctacaaa aactcagagc aaagggtatc cggaacccag accactcgcg ggcactgatc
     1921 aaggagaaat tgactgctga ccctgacagt gaggtggcca ctacaagtct ccgggtgtca
     1981 ctcatgtgcc cgctagggaa gatgcgcctg actgtccctt gtcgtgccct cacctgcgcc
     2041 cacctgcaga gcttcgatgc tgccctttat ctacagatga atgagaagaa gcctacatgg
     2101 acatgtcctg tgtgtgacaa gaaggctccc tatgaatctc ttatcattga tggtttattt
     2161 atggagattc ttagttcctg ttcagattgt gatgagatcc aattcatgga agatggatcc
     2221 tggtgcccaa tgaaacccaa gaaggaggca tctgaggttt gccccccgcc agggtatggg
     2281 ctggatggcc tccagtacag cccagtccag gggggagatc catcagagaa taagaagaag
     2341 gtcgaagtta ttgacttgac aatagaaagc tcatcagatg aggaggatct gccccctacc
     2401 aagaagcact gttctgtcac ctcagctgcc atcccggccc tacctggaag caaaggagtc
     2461 ctgacatctg gccaccagcc atcctcggtg ctaaggagcc ctgctatggg cacgttgggt
     2521 ggggatttcc tgtccagtct cccactacat gagtacccac ctgccttccc actgggagcc
     2581 gacatccaag gtttagattt attttcattt cttcagacag agagtcagca ctatggcccc
     2641 tctgtcatca cctcactaga tgaacaggat gcccttggcc acttcttcca gtaccgaggg
     2701 accccttctc actttctggg cccactggcc cccacgctgg ggagctccca ctgcagcgcc
     2761 actccggcgc cccctcctgg ccgtgtcagc agcattgtgg cccctggggg ggccttgagg
     2821 gaggggcatg gaggacccct gccctcaggt ccctctttga ctggctgtcg gtcagacatc
     2881 atttccctgg actgagaatt ctgcagatat ccagcacagt ggcggccgct cgagtctaga
     2941 gggcccgttt aaacccgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt
     3001 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
     3061 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt
     3121 ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat
     3181 gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag ggggtatccc
     3241 cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     3301 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     3361 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     3421 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     3481 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     3541 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     3601 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     3661 aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
     3721 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
     3781 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     3841 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     3901 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg
     3961 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc
     4021 cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga tcgtttcgca
     4081 tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg
     4141 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag
     4201 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc
     4261 aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc
     4321 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg
     4381 atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc
     4441 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca
     4501 tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag
     4561 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg
     4621 gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg
     4681 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca
     4741 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc
     4801 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg
     4861 acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct
     4921 gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
     4981 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
     5041 ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     5101 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     5161 tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc
     5221 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     5281 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     5341 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     5401 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     5461 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     5521 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     5581 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     5641 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     5701 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     5761 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     5821 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     5881 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     5941 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     6001 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     6061 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     6121 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtttt tttgtttgca agcagcagat
     6181 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     6241 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     6301 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     6361 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     6421 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     6481 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     6541 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     6601 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     6661 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     6721 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     6781 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     6841 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     6901 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     6961 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     7021 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     7081 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     7141 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     7201 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     7261 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     7321 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tc
//
