LOCUS       V036003                 7537 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V036003
VERSION     V036003
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        202..581
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        582..785
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        830..848
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             931..3165
                     /label="RPS6KA6(NM_001330512)"
                     /note="RPS6KA6(NM_001330512)"
                     /gene="RPS6KA6"
     CDS             3172..3198
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             3202..3228
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             3232..3258
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     polyA_signal    3299..3523
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3569..3997
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4011..4340
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4407..5006
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /note="confers resistance to puromycin"
                     /product="puromycin N-acetyltransferase"
                     /transl_table=1
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA*"
                     /label="PuroR"
     polyA_signal    5180..5301
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5350..5366)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5374..5390
                     /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(5398..5428)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5443..5464
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5752..6337)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6508..7368)
                     /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(7369..7473)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
       61 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
      121 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg
      181 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta
      241 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      301 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      361 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      421 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
      481 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
      541 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      601 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      661 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
      721 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      781 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc
      841 actataggga gacccaagct ggctagcgtt taaacttaag cttggtaccg agctcggatc
      901 cactagtcca gtgtggtgga attcgccacc atgggtctct ctacctcagc aatttggaaa
      961 aatactcgag tggaaattgt taatccctat gaagtaaaac gaaaggtgaa ggtaaatggt
     1021 cttaaaatgg ttgatgagcc aatggaagag ggagaagcag attcttgtca tgatgaagga
     1081 gttgttaaag aaatccctat tactcatcat gttaaggaag gctatgagaa agcagatcct
     1141 gcacagtttg agttgctcaa ggttcttggt caggggtcat ttggaaaggt ttttcttgtt
     1201 agaaagaaga ccggtcctga tgctgggcag ctctatgcaa tgaaggtgtt aaaaaaagcc
     1261 tctttaaaag ttcgagacag agttcggaca aagatggaga gggatatact ggtggaagta
     1321 aatcatccat ttattgtcaa attgcactat gcctttcaga ctgaagggaa actgtactta
     1381 atactggatt ttctcagggg aggagatgtt ttcacaagat tatccaaaga ggttctgttt
     1441 acagaggaag atgtgaaatt ctacctcgca gaactggccc ttgctttgga tcatctgcac
     1501 caattaggaa ttgtttatag agacctgaag ccagaaaaca ttttgcttga tgaaatagga
     1561 catatcaaat taacagattt tggactcagc aaggagtcag tagatcaaga aaagaaggct
     1621 tactcatttt gtggtacagt agagtatatg gctcctgaag tagtaaatag gagaggccat
     1681 tcccagagtg ctgattggtg gtcatatggt gttcttatgt ttgaaatgct tactggtact
     1741 ctgccatttc aaggtaaaga cagaaatgag accatgaata tgatattaaa agcaaaactt
     1801 ggaatgcctc aatttcttag tgctgaagca caaagtcttc taaggatgtt attcaaaagg
     1861 aatccagcaa atagattggg atcagaagga gttgaagaaa tcaaaagaca tctgtttttt
     1921 gcaaatattg actgggataa attatataaa agagaagttc aacctccttt caaacctgct
     1981 tctggaaaac cagatgatac tttttgtttt gatcctgaat ttactgcaaa aacacctaaa
     2041 gattctcccg gtttgccagc cagtgcaaat gctcatcagc tcttcaaagg attcagcttt
     2101 gttgcaactt ctattgcaga agaatataaa atcactccta tcacaagtgc aaatgtatta
     2161 ccaattgttc agataaatgg aaatgctgca caatttggtg aagtatatga attgaaggag
     2221 gatattggtg ttggctccta ctctgtttgc aagcgatgca tacatgcaac taccaacatg
     2281 gaatttgcag tgaagatcat tgacaaaagt aagcgagacc cttcagaaga gattgaaata
     2341 ttgatgcgct atggacaaca tcccaacatt attactttga aggatgtctt tgatgatggt
     2401 agatatgttt accttgttac ggatttaatg aaaggaggag agttacttga ccgtattctc
     2461 aaacaaaaat gtttctcgga acgggaggct agtgatatac tatatgtaat aagtaagaca
     2521 gttgactatc ttcattgtca aggagttgtt catcgtgatc ttaaacctag taatatttta
     2581 tacatggatg aatcagccag tgcagattca atcaggatat gtgattttgg gtttgcaaaa
     2641 caacttcgag gagaaaatgg acttctctta actccatgct acactgcaaa ctttgttgca
     2701 cctgaggttc ttatgcaaca gggatatgat gctgcttgtg atatctggag tttaggagtc
     2761 cttttttaca caatgttggc tggctacact ccatttgcta atggccccaa tgatactcct
     2821 gaagagatac tgctgcgtat aggcaatgga aaattctctt tgagtggtgg aaactgggac
     2881 aatatttcag acggagcaaa ggatttgctt tcccatatgc ttcatatgga cccacatcag
     2941 cggtatactg ctgaacaaat attaaagcac tcatggataa ctcacagaga ccagttgcca
     3001 aatgatcagc caaagagaaa tgatgtgtca catgttgtta agggagcaat ggttgcaaca
     3061 tactctgccc tgactcacaa gacctttcaa ccagtcctag agcctgtagc tgcttcaagc
     3121 ttagcccagc gacggagcat gaaaaagcga acatcaactg gcctgctcga gtacccatac
     3181 gacgtacctg attatgcaac ttatccttat gatgtacctg attatgctag ctatccttat
     3241 gatgtcccag actacgcata gtctagaggg cccgtttaaa cccgctgatc agcctcgact
     3301 gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg
     3361 gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg
     3421 agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg
     3481 gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga
     3541 accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg
     3601 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     3661 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     3721 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     3781 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     3841 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     3901 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     3961 aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt
     4021 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     4081 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     4141 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     4201 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     4261 cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg
     4321 gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc
     4381 aagagacagg atgaggatcg tttcgcatga ccgagtacaa gcccacggtg cgcctcgcca
     4441 cccgcgacga cgtcccccgg gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg
     4501 ccacgcgcca caccgtcgac ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac
     4561 tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg
     4621 cggtggcggt ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg
     4681 gcccgcgcat ggccgagttg agcggttccc ggctggccgc gcagcaacag atggaaggcc
     4741 tcctggcgcc gcaccggccc aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc
     4801 ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg
     4861 agcgcgccgg ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc cccttctacg
     4921 agcggctcgg cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt
     4981 gcatgacccg caagcccggt gcctgagcgg gactctgggg ttcgaaatga ccgaccaagc
     5041 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg
     5101 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     5161 ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa
     5221 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     5281 caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc
     5341 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     5401 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     5461 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     5521 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     5581 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     5641 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     5701 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     5761 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     5821 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     5881 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     5941 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     6001 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     6061 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     6121 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     6181 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     6241 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtt tttttgtttg
     6301 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     6361 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     6421 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     6481 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     6541 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     6601 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     6661 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     6721 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     6781 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     6841 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     6901 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     6961 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     7021 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     7081 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     7141 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     7201 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     7261 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     7321 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     7381 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     7441 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     7501 cgtcgacgga tcgggagatc tcccgatccc ctatggt
//
