LOCUS       V023277                 7411 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V023277
VERSION     V023277
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..2931
                     /label="WBP11(NM_016312)"
                     /note="WBP11(NM_016312)"
                     /gene="WBP11"
     polyA_signal    3011..3235
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3281..3709
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3723..4052
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4119..4913
                     /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    5087..5208
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5257..5273)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5281..5297
                     /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(5305..5335)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5350..5371
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5659..6244)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6415..7275)
                     /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(7276..7380)
                     /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 gggacggaga
     1021 tctacatcat ccaccaagag tggaaaattt atgaacccca cagaccaagc ccgaaaggaa
     1081 gcccggaaga gagaattaaa gaagaacaaa aaacagcgca tgatggttcg agctgcagtt
     1141 ttaaagatga aggatccaaa acagataatc cgagacatgg agaaattgga tgaaatggag
     1201 tttaacccag tgcaacagcc acaattaaat gagaaagtac tgaaagacaa gcgtaaaaag
     1261 ctgcgtgaaa cctttgaacg tattctacga ctctatgaaa aagagaatcc agatatttac
     1321 aaagaattga gaaagctaga agtagaatat gaacagaaga gggctcaact tagccaatat
     1381 tttgatgctg tcaagaatgc tcagcatgtg gaagtggaga gtattccttt gccagatatg
     1441 ccacatgctc cttccaacat tttgatccag gacattccac ttcctggtgc ccagccaccc
     1501 tctatcctaa agaaaacctc agcctatgga cctccaactc gggcagtttc tatccttcct
     1561 cttcttggac atggtgttcc acgtttgccc cctggcagaa aacctcctgg ccctccccct
     1621 ggtccacctc ctcctcaagt cgtgcagatg tatggccgta aagtgggttt tgccctagat
     1681 cttccccctc gtaggcgaga tgaagacatg ttatatagtc ctgaacttgc ccagcgaggt
     1741 catgatgatg atgtttctag caccagtgaa gatgatggct atcctgagga catggatcaa
     1801 gataagcatg atgacagtac tgatgacagt gacaccgaca aatcagatgg agaaagtgac
     1861 ggggatgaat ttgtgcaccg tgataatggt gagagagaca acaatgaaga aaagaagtca
     1921 ggtctgagtg tacggtttgc agatatgcct ggaaaatcaa ggaagaaaaa gaagaacatg
     1981 aaggaactga ctcctcttca agccatgatg cttcgtatgg caggtcaaga aatccctgag
     2041 gagggacggg aagtagagga attttcagag gacgatgatg aagatgattc tgatgactct
     2101 gaagcagaaa agcaatcaca aaagcagcat aaagaggaat cccattctga tggcacatcc
     2161 actgcttctt cacagcagca ggctccgccg cagtctgttc ctccttctca gatacaagca
     2221 cctcccatgc caggaccacc acctcttgga ccaccacctg ctccaccatt acggcctcct
     2281 gggccaccta caggccttcc tcctggtcca cctccaggag ctcctccatt cctgagacca
     2341 cctggaatgc caggactccg agggccctta ccccgacttt tacctccagg accaccacca
     2401 ggccgacccc ctggccctcc cccaggtcca cctccaggtc tgcctcctgg tccccctcct
     2461 cgtggacccc caccaaggct acctccccct gcacctccag gtattcctcc acctcgtcct
     2521 ggcatgatgc gcccaccttt ggtgcctccc cttggacctg ccccccctgg gctgttccca
     2581 ccagctccct tgccaaaccc tggggtttta agtgccccac ccaacttgat tcagcgaccc
     2641 aaggcggatg atacaagtgc agccaccatt gagaagaaag ccacagcaac catcagtgcc
     2701 aagccacaga tcactaatcc caaggcagag attactcgat ttgtgcccac tgcactgaga
     2761 gtacgtcggg agaataaagg ggctactgct gctccccaaa gaaagtcaga ggatgattct
     2821 gctgtgcctc ttgccaaagc agcacccaaa tctggtcctt ctgttcctgt ctcagtacaa
     2881 actaaggatg atgtctatga ggctttcatg aaagagatgg aagggctact gtgagaattc
     2941 tgcagatatc cagcacagtg gcggccgctc gagtctagag ggcccgttta aacccgctga
     3001 tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct
     3061 tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca
     3121 tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag
     3181 ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct
     3241 gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca
     3301 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     3361 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     3421 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     3481 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     3541 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     3601 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     3661 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga
     3721 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     3781 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     3841 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     3901 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     3961 tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag
     4021 gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt
     4081 tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc
     4141 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     4201 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     4261 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat
     4321 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     4381 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
     4441 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     4501 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     4561 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     4621 ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc
     4681 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     4741 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     4801 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     4861 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac
     4921 tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     4981 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     5041 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc
     5101 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     5161 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat
     5221 accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     5281 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     5341 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     5401 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     5461 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     5521 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     5581 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     5641 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     5701 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     5761 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     5821 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     5881 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     5941 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     6001 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     6061 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     6121 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     6181 caccgctggt agcggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     6241 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     6301 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     6361 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     6421 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     6481 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     6541 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     6601 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     6661 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     6721 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     6781 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     6841 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     6901 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     6961 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     7021 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     7081 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     7141 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     7201 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     7261 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     7321 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     7381 cacatttccc cgaaaagtgc cacctgacgt c
//
