LOCUS       V021034                 6765 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V021034
VERSION     V021034
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             991..2286
                     /label="PAG1(NM_018440)"
                     /note="PAG1(NM_018440)"
                     /gene="PAG1"
     polyA_signal    2365..2589
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2635..3063
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3077..3406
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3473..4267
                     /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    4441..4562
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4611..4627)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4635..4651
                     /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(4659..4689)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4704..4725
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5013..5598)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5769..6629)
                     /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(6630..6734)
                     /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 atggggcccg cggggagcct gctgggcagc
     1021 ggacagatgc agatcaccct gtggggaagt ctggctgctg tcgccatttt cttcgtcatc
     1081 accttcctca tcttcctgtg ctctagttgt gacagggaaa agaagccgcg acagcatagt
     1141 ggggaccatg agaacctgat gaacgtgcct tcagacaagg agatgttcag ccgttcagtt
     1201 actagcctgg caacagatgc tcctgccagc agtgagcaga atggggcact caccaatggg
     1261 gacattcttt cagaggacag tactctgacc tgcatgcagc attacgagga agtccagaca
     1321 tcggcctcgg atctgctgga ttcccaggac agcacaggga aaccaaaatg tcatcagagt
     1381 cgggagctgc ccagaatccc tcccgagagc gcagtggata ccatgctcac ggcgagaagt
     1441 gtggacgggg accaggggct ggggatggaa gggccctatg aagtgctcaa ggacagctcc
     1501 tcccaagaaa acatggtgga ggactgcttg tatgaaactg tgaaagagat caaggaggtg
     1561 gctgcagctg cacacctgga gaaaggccac agtggcaagg caaaatctac ttctgcctcg
     1621 aaagagctcc cagggcccca gactgaaggc aaagctgagt ttgctgaata tgcctcggtg
     1681 gacagaaaca aaaaatgtcg tcaaagtgtt aatgtagaga gtatccttgg aaattcatgt
     1741 gatccagaag aggaggcccc accacctgtc cctgttaagc ttctggacga gaatgaaaac
     1801 cttcaggaga aggaaggggg agaggcggaa gagagtgcca cagacacgac cagtgaaact
     1861 aacaagagat ttagctcatt gtcatacaag tctcgggaag aagaccccac tctcacagaa
     1921 gaagagatct cagctatgta ctcatcagta aataaacctg gacagttagt gaataaatcg
     1981 gggcagtcgc ttacagttcc ggagtccacc tacacctcca ttcaagggga cccacagagg
     2041 tcaccctcct cctgtaatga tctctatgct actgttaaag acttcgaaaa aactccaaac
     2101 agcacacttc caccagcagg gaggcccagc gaggagccag agcctgatta tgaagcgata
     2161 cagactctca acagagagga agaaaaggcc accctgggga ccaatggcca ccacggtctc
     2221 gtcccaaagg agaacgacta cgagagcata agtgacttgc agcaaggcag agatattacc
     2281 aggctctaga attctgcaga tatccagcac agtggcggcc gctcgagtct agagggcccg
     2341 tttaaacccg ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc
     2401 cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa
     2461 atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg
     2521 ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg
     2581 gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc
     2641 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     2701 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     2761 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     2821 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
     2881 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     2941 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
     3001 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     3061 attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg
     3121 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg
     3181 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc
     3241 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     3301 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt
     3361 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc
     3421 ttgtatatcc attttcggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga
     3481 acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga
     3541 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg
     3601 gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga
     3661 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt
     3721 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct
     3781 gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct
     3841 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg
     3901 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca
     3961 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga
     4021 tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt
     4081 ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt
     4141 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct
     4201 ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt
     4261 cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca
     4321 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg
     4381 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc
     4441 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     4501 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     4561 tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg
     4621 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     4681 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     4741 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     4801 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     4861 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     4921 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     4981 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     5041 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     5101 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     5161 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     5221 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     5281 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     5341 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     5401 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     5461 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     5521 tccggcaaac aaaccaccgc tggtagcggt ttttttgttt gcaagcagca gattacgcgc
     5581 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     5641 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     5701 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     5761 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     5821 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     5881 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     5941 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     6001 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     6061 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     6121 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     6181 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     6241 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     6301 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     6361 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     6421 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     6481 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     6541 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     6601 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     6661 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     6721 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtc
//
