LOCUS       V032334                 6947 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V032334
VERSION     V032334
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             908..2566
                     /label="ATP5F1A(NM_001001937)"
                     /note="ATP5F1A(NM_001001937)"
                     /gene="ATP5F1A"
     CDS             2573..2662
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     polyA_signal    2709..2933
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2979..3407
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3421..3750
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3817..4416
                     /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    4590..4711
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4760..4776)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4784..4800
                     /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(4808..4838)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4853..4874
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5162..5747)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5918..6778)
                     /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(6779..6883)
                     /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 cgccaccatg ctgtccgtgc gcgttgctgc ggccgtggtc cgcgcccttc ctcggcgggc
      961 cggactggtc tccagaaatg ctttgggttc atctttcatt gctgcaagga acttccatgc
     1021 ctctaacact catcttcaaa agactgggac tgctgagatg tcctctattc ttgaagagcg
     1081 tattcttgga gctgatacct ctgttgatct tgaagaaact gggcgtgtct taagtattgg
     1141 tgatggtatt gcccgcgtac atgggctgag gaatgttcaa gcagaagaaa tggtagagtt
     1201 ttcttcaggc ttaaagggta tgtccttgaa cttggaacct gacaatgttg gtgttgtcgt
     1261 gtttggaaat gataaactaa ttaaggaagg agatatagtg aagaggacag gagccattgt
     1321 ggacgttcca gttggtgagg agctgttggg tcgtgtagtt gatgcccttg gtaatgctat
     1381 tgatggaaag ggtccaattg gttccaagac gcgtaggcga gttggtctga aagcccccgg
     1441 tatcattcct cgaatttcag tgcgggaacc aatgcagact ggcattaagg ctgtggatag
     1501 cttggtgcca attggtcgtg gtcagcgtga actgattatt ggtgaccgac agactgggaa
     1561 aacctcaatt gctattgaca caatcattaa ccagaaacgt ttcaatgatg gatctgatga
     1621 aaagaagaag ctgtactgta tttatgttgc tattggtcaa aagagatcca ctgttgccca
     1681 gttggtgaag agacttacag atgcagatgc catgaagtac accattgtgg tgtcggctac
     1741 ggcctcggat gctgccccac ttcagtacct ggctccttac tctggctgtt ccatgggaga
     1801 gtattttaga gacaatggca aacatgcttt gatcatctat gacgacttat ccaaacaggc
     1861 tgttgcttac cgtcagatgt ctctgttgct ccgccgaccc cctggtcgtg aggcctatcc
     1921 tggtgatgtg ttctacctac actcccggtt gctggagaga gcagccaaaa tgaacgatgc
     1981 ttttggtggt ggctccttga ctgctttgcc agtcatagaa acacaggctg gtgatgtgtc
     2041 tgcttacatt ccaacaaatg tcatttccat cactgacgga cagatcttct tggaaacaga
     2101 attgttctac aaaggtatcc gccctgcaat taacgttggt ctgtctgtat ctcgtgtcgg
     2161 atccgctgcc caaaccaggg ctatgaagca ggtagcaggt accatgaagc tggaattggc
     2221 tcagtatcgt gaggttgctg cttttgccca gttcggttct gacctcgatg ctgccactca
     2281 acaacttttg agtcgtggcg tgcgtctaac tgagttgctg aagcaaggac agtattctcc
     2341 catggctatt gaagaacaag tggctgttat ctatgcgggt gtaaggggat atcttgataa
     2401 actggagccc agcaagatta caaagtttga gaatgctttc ttgtctcatg tcgtcagcca
     2461 gcaccaagcc ttgttgggca ctatcagggc tgatggaaag atctcagaac aatcagatgc
     2521 aaagctgaaa gagattgtaa caaatttctt ggctggattt gaagctgata tcgaacaaaa
     2581 actaatatca gaggaagatt tggaacagaa actaatcagt gaagaagatt tagagcaaaa
     2641 gttaatttcc gaggaggact tatagctcga gtctagaggg cccgtttaaa cccgctgatc
     2701 agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc
     2761 cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc
     2821 gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg
     2881 ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga
     2941 ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt
     3001 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     3061 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     3121 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     3181 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     3241 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     3301 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc
     3361 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat
     3421 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     3481 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
     3541 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     3601 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     3661 tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga
     3721 ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc
     3781 ggatctgatc aagagacagg atgaggatcg tttcgcatga ccgagtacaa gcccacggtg
     3841 cgcctcgcca cccgcgacga cgtcccccgg gccgtacgca ccctcgccgc cgcgttcgcc
     3901 gactaccccg ccacgcgcca caccgtcgac ccggaccgcc acatcgagcg ggtcaccgag
     3961 ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac
     4021 gacggcgccg cggtggcggt ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc
     4081 gccgagatcg gcccgcgcat ggccgagttg agcggttccc ggctggccgc gcagcaacag
     4141 atggaaggcc tcctggcgcc gcaccggccc aaggagcccg cgtggttcct ggccaccgtc
     4201 ggcgtctcgc ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg
     4261 gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc
     4321 cccttctacg agcggctcgg cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg
     4381 cgcacctggt gcatgacccg caagcccggt gcctgagcgg gactctgggg ttcgaaatga
     4441 ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg
     4501 aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg
     4561 atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca
     4621 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     4681 gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct
     4741 agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     4801 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     4861 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     4921 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     4981 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     5041 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     5101 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     5161 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     5221 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     5281 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     5341 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     5401 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     5461 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     5521 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     5581 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     5641 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtt
     5701 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     5761 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     5821 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     5881 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     5941 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     6001 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     6061 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     6121 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     6181 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     6241 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     6301 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     6361 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     6421 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     6481 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     6541 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     6601 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     6661 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     6721 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     6781 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     6841 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     6901 tgccacctga cgtcgacgga tcgggagatc tcccgatccc ctatggt
//
