LOCUS       V021521                 6754 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V021521
VERSION     V021521
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             1033..2307
                     /label="RBBP4(NM_005610)"
                     /note="RBBP4(NM_005610)"
                     /gene="RBBP4"
     polyA_signal    2354..2578
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2624..3052
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3066..3395
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3462..4256
                     /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    4430..4551
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4600..4616)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4624..4640
                     /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(4648..4678)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4693..4714
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5002..5587)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5758..6618)
                     /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(6619..6723)
                     /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 tcggatccac tagtccagtg
     1021 tggtggaatt ctatggccga caaggaagca gccttcgacg acgcagtgga agaacgagtg
     1081 atcaacgagg aatacaaaat atggaaaaag aacacccctt ttctttatga tttggtgatg
     1141 acccatgctc tggagtggcc cagcctaact gcccagtggc ttccagatgt aaccagacca
     1201 gaagggaaag atttcagcat tcatcgactt gtcctgggga cacacacatc ggatgaacaa
     1261 aaccatcttg ttatagccag tgtgcagctc cctaatgatg atgctcagtt tgatgcgtca
     1321 cactacgaca gtgagaaagg agaatttgga ggttttggtt cagttagtgg aaaaattgaa
     1381 atagaaatca agatcaacca tgaaggagaa gtaaacaggg cccgttatat gccccagaac
     1441 ccttgtatca tcgcaacaaa gactccttcc agtgatgttc ttgtttttga ctatacaaaa
     1501 catccttcta aaccagatcc ttctggagag tgcaacccag acttgcgtct ccgtggacat
     1561 cagaaggaag gctatgggct ttcttggaac ccaaatctca gtgggcactt acttagtgct
     1621 tcagatgacc ataccatctg cctgtgggac atcagtgccg ttccaaagga gggaaaagtg
     1681 gtagatgcga agaccatctt tacagggcat acggcagtag tagaagatgt ttcctggcat
     1741 ctactccatg agtctctgtt tgggtcagtt gctgatgatc agaaacttat gatttgggat
     1801 actcgttcaa acaatacttc caaaccaagc cactcagttg atgctcacac tgctgaagtg
     1861 aactgccttt ctttcaatcc ttatagtgag ttcattcttg ccacaggatc agctgacaag
     1921 actgttgcct tgtgggatct gagaaatctg aaacttaagt tgcattcctt tgagtcacat
     1981 aaggatgaaa tattccaggt tcagtggtca cctcacaatg agactatttt agcttccagt
     2041 ggtactgatc gcagactgaa tgtctgggat ttaagtaaaa ttggagagga acaatcccca
     2101 gaagatgcag aagacgggcc accagagttg ttgtttattc atggtggtca tactgccaag
     2161 atatctgatt tctcctggaa tcccaatgaa ccttgggtga tttgttctgt atcagaagac
     2221 aatatcatgc aagtgtggca aatggcagag aacatttata atgatgaaga ccctgaagga
     2281 agcgtggatc cagaaggaca agggtcctag ctcgagtcta gagggcccgt ttaaacccgc
     2341 tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg
     2401 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt
     2461 gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc
     2521 aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct
     2581 tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc ctgtagcggc
     2641 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     2701 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     2761 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     2821 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     2881 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     2941 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
     3001 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt
     3061 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     3121 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
     3181 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
     3241 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     3301 ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt
     3361 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca
     3421 ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat
     3481 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac
     3541 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc
     3601 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc
     3661 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag
     3721 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc
     3781 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg
     3841 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc
     3901 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc
     3961 cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga
     4021 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
     4081 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg
     4141 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg
     4201 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg
     4261 gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga
     4321 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
     4381 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat
     4441 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     4501 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
     4561 tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     4621 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     4681 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     4741 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     4801 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     4861 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     4921 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     4981 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     5041 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     5101 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     5161 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     5221 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     5281 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     5341 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     5401 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     5461 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     5521 aaccaccgct ggtagcggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     5581 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     5641 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     5701 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     5761 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     5821 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     5881 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     5941 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     6001 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     6061 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     6121 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     6181 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     6241 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     6301 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     6361 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     6421 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     6481 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     6541 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     6601 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     6661 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     6721 gcgcacattt ccccgaaaag tgccacctga cgtc
//
