LOCUS       V037300                 6238 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V037300
VERSION     V037300
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             937..1989
                     /label="Prkaca(NM_008854)"
                     /note="Prkaca(NM_008854)"
                     /gene="Prkaca"
     CDS             2002..2067
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    2102..2326
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2372..2800
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        2814..3143
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3155..3754
                     /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    3914..4035
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4084..4100)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4108..4124
                     /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(4132..4162)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4177..4198
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4486..5071)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5242..6102)
                     /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(6103..6207)
                     /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 aagctttcta gagaattcga tatcggatcc gccaccatgg gcaacgccgc cgccgccaag
      961 aagggcagcg agcaggagag cgtgaaagag ttcctagcca aagccaagga agatttcctg
     1021 aaaaaatggg agaccccttc tcagaataca gcccagttgg atcagtttga tagaatcaag
     1081 acccttggca ccggctcctt tgggcgagtg atgctggtga agcacaagga gagtgggaac
     1141 cactacgcca tgaagatctt agacaagcag aaggtggtga agctaaagca gatcgagcac
     1201 actctgaatg agaagcgcat cctgcaggcc gtcaacttcc cgttcctggt caaacttgaa
     1261 ttctccttca aggacaactc aaacctgtac atggtcatgg agtatgtagc tggtggcgag
     1321 atgttctccc acctacggcg gattggaagg ttcagcgagc cccatgcccg tttctacgcg
     1381 gcgcagatcg tcctgacctt tgagtatctg cactccctgg acctcatcta ccgggacctg
     1441 aagcccgaga atcttctcat cgaccagcag ggctatattc aggtgacaga cttcggtttt
     1501 gccaagcgtg tgaaaggccg tacttggacc ttgtgtggga cccctgagta cttggccccc
     1561 gagattatcc tgagcaaagg ctacaacaag gctgtggact ggtgggctct cggagtcctc
     1621 atctacgaga tggctgctgg ttacccaccc ttcttcgctg accagcctat ccagatctat
     1681 gagaaaatcg tctctgggaa ggtgcggttc ccatcccact tcagctctga cttgaaggac
     1741 ctgctgcgga accttctgca ggtggatctc accaagcgct ttgggaacct caagaacggg
     1801 gtcaatgaca tcaagaacca caagtggttt gccacgactg actggattgc catctatcag
     1861 agaaaggtgg aagctccctt cataccaaag tttaaaggcc ctggggacac gagtaacttt
     1921 gacgactatg aggaggaaga gatccgggtc tccatcaatg agaagtgtgg caaggagttt
     1981 actgagtttc tcgagaccgg tgactacaaa gaccatgacg gtgattataa agatcatgac
     2041 atcgattaca aggatgacga tgacaagtga ccggtaagtt taaaccgctg atcagcctcg
     2101 actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc
     2161 ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt
     2221 ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat
     2281 tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa
     2341 agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg
     2401 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     2461 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     2521 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     2581 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     2641 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     2701 aaccctatct cggtctattc ttttgattta taagggattt tggccatttc ggcctattgg
     2761 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc
     2821 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     2881 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     2941 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     3001 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     3061 atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt
     3121 ttggaggcct aggcttttgc aaaaagctcc cgggatgacc gagtacaagc ccacggtgcg
     3181 cctcgccacc cgcgacgacg tccccagggc cgtacgcacc ctcgccgccg cgttcgccga
     3241 ctaccccgcc acgcgccaca ccgtcgatcc ggaccgccac atcgagcggg tcaccgagct
     3301 gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga
     3361 cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc
     3421 cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc agcaacagat
     3481 ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg
     3541 cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga
     3601 ggcggccgag cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc gcaacctccc
     3661 cttctacgag cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg
     3721 cacctggtgc atgacccgca agcccggtgc ctgattcgaa atgaccgacc aagcgacgcc
     3781 caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg
     3841 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt
     3901 cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     3961 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     4021 catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc
     4081 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     4141 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     4201 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     4261 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     4321 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     4381 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     4441 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     4501 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     4561 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     4621 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     4681 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     4741 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     4801 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     4861 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     4921 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     4981 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtttttttg tttgcaagca
     5041 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     5101 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     5161 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     5221 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     5281 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     5341 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     5401 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     5461 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     5521 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     5581 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     5641 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     5701 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     5761 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     5821 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     5881 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     5941 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     6001 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     6061 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     6121 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     6181 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtc
//
