LOCUS       V040499                 7633 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V040499
VERSION     V040499
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"
     CDS             964..3123
                     /label="CNKSR1(NM_001297647)"
                     /note="CNKSR1(NM_001297647)"
                     /gene="CNKSR1"
     CDS             3130..3195
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    3233..3457
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3503..3931
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3945..4274
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4341..5135
                     /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    5309..5430
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5479..5495)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5503..5519
                     /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(5527..5557)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5572..5593
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5881..6466)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6637..7497)
                     /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(7498..7602)
                     /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 aagcttggta ccgagctcgg atccactagt ccagtgtggt ggaattcgcc
      961 accatggaac cggtagagac ctggaccccc ggaaaggtgg caacttggct gagaggtctt
     1021 gacgactccc tgcaggacta tccctttgag gactggcagc tgcctggcaa gaacctgctc
     1081 cagctctgcc cccaaagcct cgaggctctg gctgtgcggt ctctgggaca ccaggagctc
     1141 atcctgggcg gggtggaaca gctccaggcc ctgagctcca ggctacagac agagaacctg
     1201 caaagcctga cagagggact tctgggggca acccatgact tccagagcat agtccaaggc
     1261 tgcctggggg actgtgccaa gacccctatt gatgtcctct gtgcagctgt ggagctgttg
     1321 catgaagctg acgccctcct cttctggctc agcaggtacc tcttctccca cttaaatgat
     1381 ttctcagcat gccaggagat ccgagacttg ttggaggagc tgagccaggt cttgcatgag
     1441 gatggtccag cggctgagaa ggagggcaca gtcctgagga tctgcagcca cgtggctggg
     1501 atctgccaca acatcctggt ctgctgcccc aaggagctgc tggaacagaa ggccgtgctc
     1561 gagcaggtgc agctggacag tccattgggc ctagaaattc acaccaccag caattgccag
     1621 cactttgtgt cccaagtgga cacccaggtt cccactgact cccgactgca gatccagcct
     1681 ggagacgagg ttgtccagat caacgagcag gtggtggtgc gtgaggagag ggacatggtg
     1741 ggatggcccc gtaagaacat ggtgagggaa ctgctgcggg agccagccgg actcagctta
     1801 gtgctgaaga agatcccgat accggagacc cccccacaga cgccccctca ggtcctggac
     1861 tccccgcacc agaggagccc atcactgtct ctggccccac tgtctcccag ggccccatct
     1921 gaagacgtct ttgcctttga cctgtcttca aaccccagtc ccggacccag ccctgcctgg
     1981 acagactctg cctcccttgg ccctgagccc ctgcccatcc ccccggaacc cccagccata
     2041 ctcccagcag gggtagcagg gactccaggg ctccctgaat cccctgacaa gagtcctgtt
     2101 ggtcggaaga aatcaaaagg cctggcgacc cggctgagcc gccggcgggt gtcatgccgt
     2161 gagctgggcc ggccggactg tgacggctgg ctcctgttgc gaaaggcacc gggcggcttc
     2221 atgggcccgc gctggcgccg ccgctggttt gtgctcaagg gacacacgct ctactggtac
     2281 cgccagcccc aggatgagaa ggctgagggc ctcatcaatg tctccaacta tagtctggaa
     2341 agtggacatg atcagaagaa gaaatatgtg tttcagctca cccatgatgt gtacaaaccc
     2401 ttcatcttcg ctgctgatac cctgacagat ctgagcatgt gggtgcgtca tctcattacc
     2461 tgcatctcca agtaccagtc tccaggccgg gcccccccac cccgagagga agactgctac
     2521 agtgagaccg aagcagagga cccggacgat gaggctgggt cccactcagc ctcgcccagc
     2581 cctgctcaag ctgggagtcc cctccatgga gacacatcac ctgcagccac ccccacacag
     2641 cgcagcccac ggacctcctt tggctctctg acagacagca gtgaagaggc actggaagga
     2701 atggtacggg ggctgaggca gggtggcgtg tccctcctag gccagccaca gcccctgacc
     2761 caggaacagt ggcggagctc tttcatgcgg cgcaaccgag accctcagct caatgagcga
     2821 gtgcaccgtg tgcgggcgct acagagcaca ctcaaggcaa agctgcagga gctgcaggtc
     2881 ctagaagaag tgctgggtga ccctgagctg acaggagaga agttccgcca gtggaaggag
     2941 cagaaccggg agctgtactc agagggcctg ggggcctggg gagtggcaca ggctgaaggc
     3001 agctcccaca tcttgacctc tgactccaca gaacagtccc cccactccct gccctctgac
     3061 cctgaagagc actcccatct ctgccccctg acctcagaga gcagcctccg acctcctgac
     3121 ctcctcgagg actacaaaga ccatgacggt gattataaag atcatgacat cgactacaag
     3181 gatgacgatg acaagtagtg agggcccgtt taaacccgct gatcagcctc gactgtgcct
     3241 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt
     3301 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg
     3361 tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac
     3421 aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc
     3481 tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     3541 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     3601 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     3661 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     3721 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     3781 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     3841 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     3901 gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt
     3961 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     4021 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     4081 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     4141 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     4201 ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     4261 taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct gatcaagaga
     4321 caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg
     4381 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg
     4441 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt
     4501 ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg
     4561 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat
     4621 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat
     4681 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg
     4741 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg
     4801 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc
     4861 tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc
     4921 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
     4981 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg
     5041 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
     5101 tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac
     5161 cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga
     5221 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
     5281 tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa
     5341 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     5401 tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta
     5461 gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     5521 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     5581 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     5641 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     5701 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     5761 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     5821 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     5881 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     5941 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     6001 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     6061 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     6121 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     6181 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     6241 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     6301 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     6361 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggttt
     6421 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     6481 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     6541 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     6601 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     6661 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     6721 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     6781 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     6841 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     6901 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     6961 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     7021 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     7081 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     7141 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     7201 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     7261 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     7321 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     7381 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     7441 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     7501 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     7561 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     7621 gccacctgac gtc
//
