LOCUS       V039796                 6706 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V039796
VERSION     V039796
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             925..2457
                     /label="SPAM1(NM_003117)"
                     /note="SPAM1(NM_003117)"
                     /gene="SPAM1"
     CDS             2470..2535
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    2570..2794
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2840..3268
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3282..3611
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3623..4222
                     /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    4382..4503
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4552..4568)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4576..4592
                     /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(4600..4630)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4645..4666
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4954..5539)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5710..6570)
                     /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(6571..6675)
                     /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 gagaattcgc caccatggga gtgctaaaat tcaagcacat ctttttcaga
      961 agctttgtta aatcaagtgg agtatcccag atagttttca ccttccttct gattccatgt
     1021 tgcttgactc tgaatttcag agcacctcct gttattccaa atgtgccttt cctctgggcc
     1081 tggaatgccc caagtgaatt ttgtcttgga aaatttgatg agccactaga tatgagcctc
     1141 ttctctttca taggaagccc ccgaataaac gccaccgggc aaggtgttac aatattttat
     1201 gttgatagac ttggctacta tccttacata gattcaatca caggagtaac tgtgaatgga
     1261 ggaatccccc agaagatttc cttacaagac catctggaca aagctaagaa agacattaca
     1321 ttttatatgc cagtagacaa tttgggaatg gctgttattg actgggaaga atggagaccc
     1381 acttgggcaa gaaactggaa acctaaagat gtttacaaga ataggtctat tgaattggtt
     1441 cagcaacaaa atgtacaact tagtctcaca gaggccactg agaaagcaaa acaagaattt
     1501 gaaaaggcag ggaaggattt cctggtagag actataaaat tgggaaaatt acttcggcca
     1561 aatcacttgt ggggttatta tctttttccg gattgttaca accatcacta taagaaaccc
     1621 ggttacaatg gaagttgctt caatgtagaa ataaaaagaa atgatgatct cagctggttg
     1681 tggaatgaaa gcactgctct ttacccatcc atttatttga acactcagca gtctcctgta
     1741 gctgctacac tctatgtgcg caatcgagtt cgggaagcca tcagagtttc caaaatacct
     1801 gatgcaaaaa gtccacttcc ggtttttgca tatacccgca tagtttttac tgatcaagtt
     1861 ttgaaattcc tttctcaaga tgaacttgtg tatacatttg gcgaaactgt tgctctgggt
     1921 gcttctggaa ttgtaatatg gggaaccctc agtataatgc gaagtatgaa atcttgcttg
     1981 ctcctagaca attacatgga gactatactg aatccttaca taatcaacgt cacactagca
     2041 gccaaaatgt gtagccaagt gctttgccag gagcaaggag tgtgtataag gaaaaactgg
     2101 aattcaagtg actatcttca cctcaaccca gataattttg ctattcaact tgagaaaggt
     2161 ggaaagttca cagtacgtgg aaaaccgaca cttgaagacc tggagcaatt ttctgaaaaa
     2221 ttttattgca gctgttatag caccttgagt tgtaaggaga aagctgatgt aaaagacact
     2281 gatgctgttg atgtgtgtat tgctgatggt gtctgtatag atgcttttct aaaacctccc
     2341 atggagacag aagaacctca aattttctac aatgcttcac cctccacact atctgccaca
     2401 atgttcattt ggaggctgga agtctgggat caaggtatta gcagaattgg tttcttcctc
     2461 gagaccggtg actacaaaga ccatgacggt gattataaag atcatgacat cgattacaag
     2521 gatgacgatg acaagtgacc ggtaagttta aaccgctgat cagcctcgac tgtgccttct
     2581 agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc
     2641 actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt
     2701 cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat
     2761 agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg
     2821 ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     2881 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     2941 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     3001 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     3061 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     3121 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     3181 gtctattctt ttgatttata agggattttg gccatttcgg cctattggtt aaaaaatgag
     3241 ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg
     3301 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     3361 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     3421 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     3481 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     3541 aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     3601 gcttttgcaa aaagctcccg ggatgaccga gtacaagccc acggtgcgcc tcgccacccg
     3661 cgacgacgtc cccagggccg tacgcaccct cgccgccgcg ttcgccgact accccgccac
     3721 gcgccacacc gtcgatccgg accgccacat cgagcgggtc accgagctgc aagaactctt
     3781 cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt
     3841 ggcggtctgg accacgccgg agagcgtcga agcgggggcg gtgttcgccg agatcggccc
     3901 gcgcatggcc gagttgagcg gttcccggct ggccgcgcag caacagatgg aaggcctcct
     3961 ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc accgtcggcg tctcgcccga
     4021 ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg
     4081 cgccggggtg cccgccttcc tggagacctc cgcgccccgc aacctcccct tctacgagcg
     4141 gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat
     4201 gacccgcaag cccggtgcct gattcgaaat gaccgaccaa gcgacgccca acctgccatc
     4261 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg
     4321 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc
     4381 caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     4441 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     4501 ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct
     4561 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     4621 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     4681 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     4741 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     4801 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     4861 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     4921 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     4981 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     5041 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     5101 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     5161 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     5221 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     5281 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     5341 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     5401 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     5461 atccggcaaa caaaccaccg ctggtagcgg tttttttgtt tgcaagcagc agattacgcg
     5521 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     5581 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     5641 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     5701 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     5761 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     5821 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     5881 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     5941 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     6001 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     6061 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     6121 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     6181 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     6241 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     6301 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     6361 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     6421 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     6481 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     6541 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     6601 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     6661 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtc
//
